biotechrabbit
Product Catalog
2013 2014
About biotechrabbit
Scientists around the world are working to leap ahead of diseases and epidemics and to fuel our lives with innovations. Aside from brilliant minds and relentless passion, the success of science depends on the quality of the materials used.
biotechrabbit is determined to offer the best products and innovative solutions to those who lead the progress. We seek to serve our customers with the newest life science products and services. We care deeply about our relationships with our partners and customers. It is our flexibility and our genuine concern that enable us to offer highly customized solutions for their specific requirements. We want our partners and customers to leap and lead the progress of life science.
Our team of highly engaged scientists, experienced managers and business developers is happy to be a part of the scientific community that is leading with key technologies and conducting advanced research.
Our way of making business is to combine the passion and pure curiosity of excellent researchers with the agile spirit of true entrepreneurs.
biotechrabbit leap and lead
Contact Us
biotechrabbit GmbH Neuendorfstr. 24a 16761 Hennigsdorf Germany [email protected] www.biotechrabbit.com Office: +49 3302 207541 0 Fax: +49 3302 207541 1Commercial Register: Neuruppin (HRB 9381 NP) Managing Director: Dr. Bernd Haase
Principal Office: Hennigsdorf Tax Office: Oranienburg VAT No./USt-Id: DE 279969868 Tax No.: 05310604935
We highly appreciate your contacting us personally, by telephone, fax, post or email. It is our priority to get back to you as quickly as possible.
Ordering:
Tel: +49 3302 207541 0 Fax: +49 3302 207541 1
OEM and Antibody Service Tel: +49 3302 207541 5 Fax: +49 3302 207541 1 [email protected] Technical Support: Tel: +49 3302 207541 2 Tel: +49 3302 207541 0 Fax: +4903302 207541 1 [email protected] Sales: Tel: +49 3302 207541 0 Fax: +49 3302 207541 1 [email protected]
Collaboration and Partnership: Tel: +49 3302 207541 5
Career Opportunities: Tel :+49 3302 207541 0
Table of Contents
OEM and Other Services 4
Antibody Services 5
Standard PCR Product Selection Guide 6
Taq DNA Polymerase, recombinant, 5 U/µl 7
Taq DNA Polymerase, convenient, 5 U/µl 7
PCR Master Mix, 2x 8
Green Taq DNA Polymerase, 5 U/µl 9
Green PCR Master Mix, 2x 9
Red Taq DNA Polymerase, 1 U/µl 10
Red PCR Master Mix, 2x 10
Hot Start PCR Product Selection Guide 11
Hot Start Taq DNA Polymerase, 5 U/µl 12
Hot Start PCR Master Mix, 2x 12
Red Hot Start DNA Polymerase, 5 U/µl 13
Red Hot Start PCR Master Mix, 2x 14
Green Hot Start PCR Master Mix, 2x 14
High Fidelity and Long Range PCR Product Selection Guide 15
Pfu DNA Polymerase, 2.5 U/µl 16
Pfu PCR Master Mix, 2x 16
Long And High Fidelity PCR Enzyme Mix, 2.5 U/µl 17
Long Range PCR Master Mix, 2x 17
Reverse Transcription and RT-PCR Product Selection Guide 18
One Step RT-PCR Kit 19
NEW RevertUP Reverse Transcriptase, 100 U/µl 19
MMuLV Reverse Transcriptase, 200 U/µl 20
RNAse Inhibitor, 40 U/µl 20
Quantitative Real-Time PCR Product Selection Guide 21
QPCR Green Master Mix, 2x Intercalating-dye-based qPCR mixes without reference dye 23 QPCR SyGreen Master Mix LRox and HRox, 2x Intercalating-dye-based qPCR mixes with ROX 24 QPCR Probe Master Mix, 2x Probe-based qPCR mixes without reference dye 25 QPCR Probe-R Master Mix LRox and HRox, 2x Probe-based qPCR mixes with ROX 26
NEW QRT-PCR SyGreen One Step Kits, LRox and HRox 27
NEW QRT-PCR Probe-R One Step Kits, LRox and HRox 28
Direct PCR/Crude Sample PCR 29
U/µl 29
dNTP Mixes and Sets 30
Nucleic Acid Purification 32
NEW UPzol RNA Isolation Solution 33
NEW GenUP Total RNA Mini Kit 33
NEW GenUP Plasmid Mini Kit 34
NEW GenUP gDNA Mini Kit 35
NEW GenUP Plant DNA Kit 35
NEW GenUP Bacterial gDNA Mini Kit 36
NEW GenUP Virus RNA Kit 37
NEW GenUP Virus RNA/DNA Kit 38
NEW GenUP PCR Cleanup Kit 39
NEW GenUP PCR/Gel Cleanup Kit 39
NEW GenUP Gel Extraction Kit 40
DNA Electrophoresis 41
DNA Electrophoresis Ladders and Loading Dyes 42
Ultrapure Enzymes for Molecular Biology 43
phi 29 DNA Polymerase, 10 U/µl 44
U/µl 45
T4 DNA Ligase, Rapid, 600 U/µl 46
Uracil DNA Glycosylase, 2 U/µl 47
Protein Electrophoresis 48
Protein Electrophoresis Ladders 49
Protein Purification Media 50
His-Tagged Protein Purification Media 51
Immobilized Metal Affinity Chromatography Media (IMAC) 52
Ion-Exchange Chromatography Protein Purification Media (IEX) 53
OEM and Other Services
Most of biotechrabbit molecular biology and proteomics products are available in bulk or as custom formulations. biotechrabbit welcomes all your OEM and bulk requests:
Tel: +49 3302 207541 5 Fax: +49 3302 207541 1
You will receive answers to your questions about the availability of OEM products within three working days.
We are a team of dedicated scientists, experienced managers and business developers headquartered in Germany. To be ahead of the newest technologies and achievements in science , we closely collaborate with scientists who are leading key technologies and conducting advanced research.
As a reliable OEM partner we offer the following:
Custom formulations and bulk production of most of the products from our portfolio
A choice of enzymes for standard PCR, high-fidelity PCR, reverse transcription, cloning and other applications Glycerin-free enzymes and antibodies, ready for lyophilization
Enzymes for diagnostics kit development Lyophilization service using special formulations
Development projects - cloning of proteins, expression optimization, purification scheme development Custom vialing, packaging and labeling
Flexibility and outstanding personal attitude
Production process documentation and quality certificates Continuous technical support
ion is to create continuous and sustainable value and added benefit for our customers. We want to serve our customers
With the newest and most advanced life science technologies and products With services to help co-develop new applications or diagnostic tests To achieve maximum performance in research and shortest time to results biotechrabbit
Antibody Services
We offer an exceptional quality and high capacity antibody production service, which is provided by qualified specialist who have been working in the field for more than 10 years.
biotechrabbit welcomes all antibody related requests: Tel: +49 3302 207541 5
Fax: +49 3302 207541 1
Our antibody production services include
Monoclonal antibodies Polyclonal antibodies
W o rk fl o w
Choice of animals: mouse and rat
Starting material 1 1.5 mg of your protein (or peptide) or the protein sequence
Development of hybridoma cells using the provided protein Generation of monoclonal antibodies against membrane
proteins using a special vector and proprietary cell immunization and screening technology
Binding and activity screening using ELISA, western blot, FACS or special activity tests
Production of a test sample of up to 10 mg
Large-scale production by fermentation for producing up to 1kg of antibody
Choice of animals: rabbit, guinea pig, goat, and sheep
others on request
Starting material approximately 1 mg protein Production of a polyclonal antiserum Antibody purification and labeling, when
required
Visit our homepage for additional services
D u ra ti o n
Hybridoma supernatants are ready for screening in 4 weeks Tests are completed in 4 weeks (depending on test ordered) When subcloning has been ordered, the first and second
subcloning of hybrodoma cells are completed in 4 6 weeks Production of monoclonal antibodies:
2 weeks for 10 mg scale 4 weeks for 200 mg scale Up to 8 weeks for larger amounts
Delivery of monoclonal antibodies in PBS buffer or Tris buffer , according to
The serum or the affinity purified antibodies are delivered in 4 weeks after we receive the protein Alternative immunization strategies available on
request
P
ri
c
e Variable dependent on the size of the project, the production
scale and on the panel of required tests Variable dependent on additional services
Benefits for the customer
State-of-the-art antibody production: fermentation performed by a team with 10 years of experience.
Shortest project time and a larger variety of monoclonal antibodies from which to select : our special immunization requires much shorter time than conventional methods (17 days).
Antibodies on demand: more than 1800 hybridomas have already been produced.
Highest purity: due to the use of genetically engineered protein A/G (lacking the albumin -binding domain) and cultivation in serum-free or protein-free media. After one-step purification with affinity chromatography , bovine IgGs and serum albumins cannot be detected in our monoclonal antibodies. Remaining aggregates can be quantitatively removed with preparative size -exclusion chromatography.
Endotoxin-free antibodies: cGMP-compliant documentation can be supplied for preclinical trials.
Yields of up to 500 mg of antibody per liter of culture per day : with our high-cell-density (>2 x 107 cells per ml) fermentation that uses an efficient two liter reactor systems, gram quantities can be delivered within weeks.
High antibody production capacity: from 3 mg to over a kilogram. Short turnaround times: due to high-capacity production facility.
No waiting times for customers: the antibody production process starts the day after receiving the starting materials. We are always ready for your project start with only short notice.
Test orders of 10 mg antibody available at low prices. Production process documentation and quality certificates. Continuous technical support, flexibility and personal attitude. biotechrabbit
Standard PCR Product Selection Guide
Standard PCR typically has the following parameters: Target length is 1 3 kb
GC content of the template is 40 60% Template is pure, abundant and unproblematic Very high-fidelity of amplification is not required
Unmodified Taq DNA polymerase is the most common enzyme for standard PCR. For simplified reaction setup, PCR master mixes which contain all necessary PCR reagents are off ered. Colored, high-density PCR buffers are available for direct loading of PCR products onto gels. The enzyme formulations in colored storage buffer s allow easy visualization of enzyme-containing reaction mixtures and monitoring the reaction set up process.
Product Main feature
Main PCR application Simplified setup Optimization freedom Direct gel loading Colored enzyme mixtures Fidelity Maximum product length TA cloning Taq DNA Polymerase, recombinant, 5 U/µl MgCl2 supplied in a
separate vial for
maximum flexibility Standard
Typical for Taq 3 5 kb
Taq DNA Polymerase, convenient, 5 U/µl Reaction Buffer contains optimal MgCl2 concentrationStandard
Typical for Taq 3 5 kb
PCR Master Mix, 2x
Premixed PCR reagents; just add template, primers and water.
Routine,
high-throughput
Typical for
Taq 3 5 kb
Green Taq DNA Polymerase, 5 U/µl
Green Reaction Buffer for direct loading onto gel after PCR Standard with direct gel loading
Typical for Taq 3 5 kb
Green PCR Master Mix, 2x Premixed PCR reagents; just add template, primers and water, including Green Reaction Buffer for direct loading onto gel after the PCR Routine, high-throughput with direct gel loading
Typical for Taq 3 5 kb
Red Taq DNA Polymerase, 1 U/µl
Red-colored Taq DNA Polymerase Storage Buffer for visualization of mixes containing the enzyme Standard with enzyme visualization
Typical for Taq 3 5 kb
Red PCR Master Mix, 2x Premixed PCR reagents; just add template, primers and water, including Red Buffer for visualization of mixes containing the enzyme Routine, high-throughput with enzyme visualization
Typical for Taq 3 5 kb
Taq DNA Polymerase, recombinant, 5 U/µl
CAT. # CONTENTS
BR0100101 100 U Taq DNA Polymerase, recombinant, 5 U/µl 1.8 ml 10x Reaction Buffer 1.5 ml 50 mM MgCl2
BR0100102 500 U Taq DNA Polymerase, recombinant, 5 U/µl 2 x 1.8 ml 10x Reaction Buffer 1.5 ml 50 mM MgCl2 APPLICATIONS Routine amplification up to 4 kb RT-PCR TA cloning FEATURES
For maximum flexibility, MgCl2 is supplied separately
High product yields in a wide application range
Exceptionally pure Taq DNA Polymerase for both routine and demanding PCR applications
COMPOSITION OF COMPONENTS
Taq DNA Polymerase, 5 U/µl, in Storage Buffer:
20 mM Tris-HCl (pH 8.0 at 25°C), 30 mM KCl, 0.1 mM DTT, 0.5% Tween 20, 50% (v/v) glycerol 10x Reaction Buffer: 670 mM Tris-HCl (pH 8.8 at 25°C), 166 mM (NH4)2SO4, 4.5% Triton® X-100, 2 mg/ml gelatin DESCRIPTION
biotechrabbit Taq DNA Polymerase is a first-choice thermostable DNA polymerase for all routine PCR applications, ensuring high product yields from various templates for targets up to 4 kb in size.
Taq DNA polymerase is a thermostable, highly processive 5'→3' DNA polymerase that has low 5'→3' exonuclease activity and lacks 3'→5' exonuclease (proofreading) activity. The latter allows incorporation of modified nucleotides into DNA.
The enzyme also exhibits deoxynucleotidyl transferase activity, resulting in the addition of an extra A overhang at the 3' ends of PCR products, allowing easy cloning of PCR products into vectors with T overhangs.
UNIT DEFINITION
One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol of dNTP into acid-insoluble form in 30 minutes at 72°C in the presence of 1x Reaction Buffer. QUALITY CONTROL Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Functional PCR Assay
Taq DNA Polymerase, convenient, 5 U/µl
CAT. # CONTENTS
BR0100702 100 U Taq DNA Polymerase, convenient, 5 U/µl 2 x 1.8 ml 10x Reaction Buffer with MgCl
2 APPLICATIONS Routine amplification up to 4 kb RT-PCR TA cloning FEATURES
Optimal MgCl2 concentration in the Reaction Buffer
High product yields in a wide application range
Exceptionally pure Taq DNA Polymerase for both routine and demanding PCR applications
COMPOSITION OF COMPONENTS
Taq DNA Polymerase, 5 U/µl, in Storage Buffer:
20 mM Tris-HCl, 100 mM NaCl, 1.0 mM DTT, 0.1 mM EDTA, Stabilizer, 50% glycerol, pH 7.5 at 25°C
10x Reaction Buffer with MgCl2:
100 mM Tris-HCl (pH 8.3 at 25°C), 500 mM KCl, 15 mM MgCl2
DESCRIPTION
biotechrabbit Taq DNA Polymerase is a first-choice thermostable DNA polymerase for all routine PCR applications, ensuring high product yields from various templates for targets up to 4 kb in size.
Taq DNA polymerase is a thermostable, highly processive 5'→3' DNA polymerase that has low 5'→3' exonuclease activity and lacks 3'→5' exonuclease (proofreading) activity. The latter allows incorporation of modified nucleotides into DNA.
The enzyme also exhibits deoxynucleotidyl transferase activity, resulting in the addition of an extra A overhang at the 3' ends of PCR products, allowing easy cloning of PCR products into vectors with T overhangs.
Purified from a recombinant E. coli strain carrying the Taq DNA polymerase gene from Thermus aquaticus YT-1. UNIT DEFINITION
One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol of dNTP into acid-insoluble form in 30 minutes at 72°C in the presence of the 1x Reaction Buffer.
QUALITY CONTROL
Single-Stranded Exonuclease Activity Double-Stranded Exonuclease Activity Double-Stranded Endonuclease Activity E. coli 16S rDNA Contamination Test Functional PCR Assay
PCR Master Mix, 2x
CAT. # CONTENTS
BR0100201 100 reactions (50 µl) 2 x 1.25 ml PCR Master Mix, 2x BR0100202 500 reactions (50 µl) 10 x 1.25 ml PCR Master Mix, 2x APPLICATIONS
Routine amplification up to 4 kb TA cloning
FEATURES
Exceptionally pure Taq DNA Polymerase and highest quality dNTPs and buffer in a 2x PCR Master Mix formulation High product yields and robustness in a wide application
range
COMPOSITION OF COMPONENTS PCR Master Mix, 2x:
Proprietary composition contains all necessary PCR reagents, including 3 mM MgCl2. (1x has 1.5 mM MgCl2)
DESCRIPTION
biotechrabbit PCR Master Mix is a perfect choice for fast reaction setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. It is designed for routine, high-throughput PCR amplification of 0.2–4 kb DNA targets.
The 2x Master Mix contains highly purified recombinant biotechrabbit Taq DNA polymerase, highest quality dNTPs and optimal PCR buffer reagents; thus, only template DNA, PCR primers and PCR-grade water need to be added. Taq DNA polymerase is a thermostable highly processive 5'→3' DNA polymerase that has low 5'→3' exonuclease activity and lacks 3'→5' exonuclease (proofreading) activity. The latter allows incorporation of modified nucleotides into DNA.
The enzyme also exhibits deoxynucleotidyl transferase activity that results in the addition of an extra A overhang at the 3' ends of PCR products, allowing easy cloning of PCR products into vectors with T overhangs.
QUALITY CONTROL Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay PCR Master Mix: Functional PCR Assay L 1 2 3 4 5 6
Picture 1. Performance of biotechrabbit PCR Master Mixes.
A 700 bp genomic DNA fragment was amplified using different PCR master mixes. Colored PCR master mixes show comparable performance to colorless mixes.
L 100 bp DNA Ladder, rtu (BR0800201) 1 PCR Master Mix, 2x (BR0100201) 2 Red PCR Master Mix, 2x (BR0100601) 3 Green PCR Master Mix, 2x (BR0100401) 4 Hot Start PCR Master Mix, 2x (BR0200201) 5 Red Hot Start PCR Master Mix, 2x (BR0200401) 6 Green Hot Start PCR Master Mix, 2x (BR0200501)
Green Taq DNA Polymerase, 5 U/µl
CAT. # CONTENTS
BR0100301 100 U 20 µl Taq DNA Polymerase, 5 U/µl 2 x 1.8 ml 5x Green Reaction Buffer 1.5 ml 50 mM MgCl2
BR0100302 500 U 100 µl Taq DNA Polymerase, 5 U/µl 4 x 1.8 ml 5x Green Reaction Buffer 1.5 ml 50 mM MgCl2
BR0100303 5 x 500 U 5 x 100 µl Taq DNA Polymerase, 5 U/µl 5 x 1.8 ml 5x Green Reaction Buffer 5 x 1.5 ml 50 mM MgCl2
BR0100304 10 x 500 U 10 x 100 µl Taq DNA Polymerase, 5 U/µl 10 x 1.8 ml 5x Green Reaction Buffer 10 x 1.5 ml 50 mM MgCl2
BR0100305 20 x 500 U 20 x 100 µl Taq DNA Polymerase, 5 U/µl 20 x 1.8 ml 5x Green Reaction Buffer 20 x 1.5 ml 50 mM MgCl2
APPLICATIONS
High-throughput PCR for immediate gel analysis Routine amplification up to 4 kb
TA cloning FEATURES
Green Reaction Buffer allows direct loading onto a gel immediately after PCR
High product yields and robustness
Exceptionally pure Taq DNA Polymerase for both routine and demanding PCR applications
COMPOSITION OF COMPONENTS
Taq DNA Polymerase, 5 U/µl, in Storage Buffer:
20 mM Tris-HCl (pH 8.0 at 25°C), 30 mM KCl, 0.1 mM DTT, 0.5% Tween 20, 50% (v/v) glycerol
5x Green Reaction Buffer with MgCl2:
335 mM Tris-HCl (pH 8.8 at 25 °C), 83 mM (NH4)2SO4, 2.25%
Triton®-X-100, 1 mg/ml gelatin, tracking dyes (yellow and blue)
DESCRIPTION
biotechrabbit Green Taq DNA Polymerase is a first-choice thermostable DNA polymerase for routine PCR applications, allowing direct electrophoresis without the need to add loading buffer and ensuring high product yields from various templates.
The 5x Green Taq Reaction Buffer is recommended for any amplification reaction that will be visualized by agarose gel electrophoresis and ethidium bromide staining. The Green Reaction Buffer contains two dyes (blue and yellow) that separate during electrophoresis, allowing the migration progress to be monitored. Reactions with Green Reaction Buffer have sufficient density for direct loading onto agarose gels.
The dyes absorb between 225 300 nm, making standard A260 readings to determine DNA concentration unreliable.
Taq DNA polymerase is a thermostable highly processive 5'→3' DNA polymerase that has low 5'→3' exonuclease activity and lacks 3'→5' exonuclease (proofreading) activity. The enzyme also exhibits deoxyribonucleotidyl transferase activity that results in the addition of an extra A overhang at the 3' ends of PCR products, allowing easy cloning of PCR products into vectors with T overhangs.
UNIT DEFINITION
One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol of dNTP into acid -insoluble form in 30 minutes at 74°C in the presence of the reaction buffer. QUALITY CONTROL Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Functional PCR Assay
Green PCR Master Mix, 2x
CAT. # CONTENTS
BR0100401 100 reactions (50 µl) 2 x 1.25 ml Green PCR Master Mix , 2x BR0100402 500 reactions (50 µl) 10 x 1.25 ml Green PCR Master Mix , 2x APPLICATIONS
High-throughput PCR for immediate gel analysis Routine amplification up to 4 kb
TA cloning FEATURES
Green PCR Master Mix formulation allows direct loading on to a gel immediately after PCR
Exceptionally pure Taq DNA Polymerase and highest quality dNTPs in a 2x Green PCR Master Mix
High product yields and robustness in a wide application range
COMPOSITION OF COMPONENTS Green PCR Master Mix, 2x:
Proprietary master mix composition contains all necessary PCR components, including 3 mM MgCl2 (1x has 1.5 mM
MgCl2) and blue and yellow electrophoresis dyes
DESCRIPTION
biotechrabbit Green PCR Master Mix is a perfect choice for a fast reaction setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. Additionally the special formulation allows reactions onto be loaded onto the gel directly after
amplification without a separate step for adding loading dye. PCR Master Mix contains two dyes (blue and yellow) that separate during electrophoresis, allowing the migration progress to be monitored. Reactions with the Green PCR Master Mix have sufficient density for direct loading on to agarose gels. Green Reaction Buffer also allows mixtures containing the enzyme to be identified.
The Green PCR Master Mix contains highly purified recombinant biotechrabbit Taq DNA polymerase, highest quality dNTPs and optimal PCR buffer reagents; thus, only template DNA, PCR primers and PCR-grade water need to be added. QUALITY CONTROL Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay
Green PCR Master Mix: Functional PCR Assay
Red Taq DNA Polymerase, 1 U/µl
CAT. # CONTENTS
BR0100501 100 U 100 µl Red Taq DNA Polymerase, 1 U/µl 1.8 ml 10x Reaction Buffer 1.5 ml 50 mM MgCl2
BR0100502 500 U 500 µl Red Taq DNA Polymerase, 1 U/µl 2 x 1.8 ml 10x Reaction Buffer 1.5 ml 50 mM MgCl2
BR0100503 5 x 500 U 5 x 500 µl Red 5 x 1.8 ml 10x Reaction Buffer Taq DNA Polymerase, 1 U/µl 5 x 1.5 ml 50 mM MgCl2
BR0100504 10 x 500 U 10 x 500 µl Red Taq DNA Polymerase, 1 U/µl 2 x 1.8 ml 10x Reaction Buffer 1.5 ml 50 mM MgCl2
BR0100505 20 x 500 U 10 x 500 µl Red Taq DNA Polymerase, 1 U/µl 10 x 1.8 ml 10x Reaction Buffer 10 x 1.5 ml 50 mM MgCl2
APPLICATIONS
High-throughput, simplified PCR setup with easy identification of reaction mixtures containing enzyme Routine amplification to 4 kb and TA cloning
FEATURES
Red Taq DNA Polymerase Storage Buffer allows mixtures containing the enzyme to be identified
High product yields and robustness
Exceptionally pure Taq DNA Polymerase for both routine and demanding PCR applications
COMPOSITION OF COMPONENTS
Taq DNA Polymerase, 5 U/µl, in Storage Buffer:
20 mM Tris-HCl (pH 8.0 at 25°C), 30 mM KCl, 0.1 mM DTT, 0.5% Tween 20, 50% (v/v) glycerol, red dye
10x Reaction Buffer:
670 mM Tris-HCl (pH 8.8 at 25°C), 166 mM (NH4)2SO4,
4.5% Triton® X-100, 2 mg/ml gelatin
DESCRIPTION
biotechrabbit Red Taq DNA Polymerase is a first-choice colored, thermostable DNA polymerase for simplifying PCR setup. This red-colored enzyme formulation performs in all respects like colorless Taq DNA Polymerase formulations and ensures high product yields from various templates. Additionally, reaction mixtures that contain the enzyme are easily identified.
Taq DNA polymerase is a thermostable highly processive 5'→3' DNA polymerase that has low 5'→3' exonuclease activity and lacks 3'→5' exonuclease (proofreading) activity. The enzyme also exhibits deoxynucleotidyl transferase activity that results in the addition of an extra A overhang at the 3' ends of PCR products, allowing easy cloning of PCR products into vectors with T overhangs.
UNIT DEFINITION
One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol of dNTP into acid-insoluble form in 30 minutes at 72°C in the presence of the reaction buffer. QUALITY CONTROL Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Functional PCR Assay
Red PCR Master Mix, 2x
CAT. # CONTENTS
BR0100601 100 reactions (50 µl) 2 x 1.25 ml Red PCR Master Mix, 2x BR0100602 500 reactions (50 µl) 10 x 1.25 ml Red PCR Master Mix , 2x APPLICATIONS
High throughput, simplified PCR setup with easy
identification of reaction mixtures containing the enzyme and dNTPs
Routine amplification up to 4 kb TA cloning
FEATURES
Exceptionally pure Taq DNA Polymerase and highest quality dNTPs in a 2x Red PCR Master Mix formulation
High product yields and robustness in a wide application range
COMPOSITION OF COMPONENTS Red PCR Master Mix, 2x:
Proprietary composition, contains all necessary PCR reagents, including 3 mM MgCl2 (1x has 1.5 mM MgCl2)
DESCRIPTION
biotechrabbit Red PCR Master Mix is a perfect choice for simplified reaction setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. Additionally the colored formulation provides the possibility to identify the reactions which already contain the enzyme and dNTPs.
This red-colored master mix performs in all respects like colorless PCR Master Mix and ensures high product yields from various templates. It contains highly purified
recombinant biotechrabbit Taq DNA polymerase, highest quality dNTPs and optimal PCR buffer reagents; thus, only template DNA, PCR primers and PCR-grade water need to be added. QUALITY CONTROL Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay
Red PCR Master Mix: Functional PCR Assay
Hot Start PCR Product Selection Guide
Hot Start PCR is a more sensitive technique than standard PCR that allows amplification of low-abundance targets and single-copy genes while reducing PCR background problems.
Thermophilic DNA polymerases are unfortunately active at room temperature, which can result in amplification of unspecific targets due to random primer annealing events. Hot-start enzymes are completely inactive during room-temperature reaction setup and become active only after heating. Thus, misprimed amplification products and primer–dimer formation do not occur, diminishing PCR background.
Hot-start enzymes are typically created with one of the following techniques: chemically, by the addition of thermo-labile blocking groups, or with binding by one or more Taq-specific antibodies. Both of these techniques prevent enzyme activity until the blockage is disrupted at high temperature.
Usually hot-start modification does not affect PCR amplification: after thermal reactivation, hot-start Taq DNA polymerases perform similarly to unmodified Taq with respect to processivity, fidelity and deoxyribonucleotidyl transferase activity, allowing TA cloning. Hot-start PCR master mixes containing all necessary PCR reagents are offered for simplified reaction setup. Colored hot-start PCR buffers are available for direct loading of PCR products onto gels.
Product Main feature
Main PCR application Simplified setup Optimization freedom Direct gel loading Colored enzyme mixtures Fidelity Maximum product length TA cloning
Hot Start Taq DNA Polymerase, 5 U/µl
Antibody-based Hot Start Taq DNA Polymerase supplied with PCR Booster for higher PCR specificity and better yield
High-sensitivity,
hot-start
Typical for Taq 3 5 kb
Hot Start PCR Master Mix, 2x
Antibody-based Hot Start Taq DNA Polymerase with all PCR reagents High-throughput, high-specificity, hot-start
Typical for Taq 3 5 kb
Red Hot Start DNA Polymerase, 5 U/µl
Antibody-based Hot Start Taq DNA Polymerase in a red-colored Storage Buffer for easy visualization of mixes containing the enzyme
High-sensitivity, hot-start with easy identification of reactions containing enzyme
Typical forTaq 3 5 kb
Red Hot Start PCR Master Mix, 2x
Antibody-based Hot Start Taq DNA Polymerase and all PCR reagents in Red Buffer for easy visualization of mixes containing the enzyme
High-throughput, high-specificity hot-start with easy identification of reactions containing enzyme
Typical for Taq 3 5 kb
Green Hot Start PCR Master Mix, 2x Antibody-based Hot Start Taq DNA Polymerase and all PCR reagents in Green Reaction Buffer for direct loading onto gels after the PCRHigh-throughput, high-specificity, hot-start for direct
loading on gels
Typical for
Hot Start Taq DNA Polymerase, 5 U/µl
CAT. # CONTENTS
BR0200101 100 U
20 µl Hot Start Taq DNA Polymerase, 5 U/µl 1.8 ml 10x Reaction Buffer
1.5 ml 50 mM MgCl2
1.8 ml 5x PCR Booster BR0200102 500 U
100 µl Hot Start Taq DNA Polymerase, 5 U/µl 2 x 1.8 ml 10x Reaction Buffer
1.5 ml 50 mM MgCl2
2 x 1.8 ml 5x PCR Booster APPLICATIONS
High-specificity, hot-start amplification up to 4 kb Amplification of low-copy-number targets RT-PCR
TA cloning FEATURES
High PCR specificity and sensitivity
Hot Start Taq DNA Polymerase for demanding, sensitive PCR applications and high yields
COMPOSITION OF COMPONENTS
Taq DNA Polymerase with bound antibody, in proprietary Storage Buffer
10x Reaction Buffer:
670 mM Tris-HCl (pH 8.8 at 25°C), 166 mM (NH4)2SO4,
4.5% Triton® X-100, 2 mg/ml gelatin
DESCRIPTION
biotechrabbit Hot Start Taq DNA polymerase is a first-choice hot-start PCR enzyme for all demanding PCR applications. The enzyme ensures high product yields with low
background; without primer–dimer formation and nonspecific priming.
The Hot Start Taq Polymerase is inactive during reaction setup due to the bound antibody which is quickly released at elevated temperatures, ensuring the enzyme is active only during PCR. There is no need for prolonged heating or denaturation steps.
The PCR specificity can be further increased in some cases by addition of the supplied 5x PCR Booster. The optional use of 5x PCR Booster improves PCR results in many cases, including impure template or low template amounts. Taq DNA polymerase is a thermostable highly processive 5'→3' DNA polymerase that has low 5'→3' exonuclease activity and lacks 3'→5' exonuclease (proofreading) activity. The enzyme also exhibits deoxynucleotidyl transferase activity that results in the addition of an extra A overhang at the 3' ends of PCR products allowing easy cloning of PCR products into vectors with T overhangs .
UNIT DEFINITION
One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol of dNTP into acid -insoluble form in 30 minutes at 72°C in the presence of the reaction buffer. QUALITY CONTROL Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Functional PCR Assay
Hot Start PCR Master Mix, 2x
CAT. # CONTENTS
BR0200201 100 reactions (50 µl) 2 x 1.25 ml Hot Start PCR Master Mix, 2x BR0200202 500 reactions (50 µl) 10 x 1.25 ml Hot Start PCR Master Mix, 2x APPLICATIONS
High-specificity, high-throughput, hot-start amplification up to 4 kb
Amplification of low-copy-number targets TA cloning
FEATURES
Highest PCR specificity and sensitivity without prolonged reactivation
Exceptionally pure Hot Start Taq DNA Polymerase and highest quality dNTPs in a 2x Hot Start PCR Master Mix COMPOSITION OF COMPONENTS
Hot Start PCR Master Mix, 2x:
Proprietary composition, contains all necessary hot-start PCR reagents, including 3 mM MgCl2. (1x has 1.5 mM MgCl2)
DESCRIPTION
biotechrabbit Hot Start PCR Master Mix is a perfect choice for a fast reaction setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. It is designed for low-background, high-throughput PCR of 0.2 5 kb DNA targets.
The 2x Hot Start PCR Master Mix contains pure
biotechrabbit Hot Start Taq DNA polymerase, highest quality dNTPs and optimal PCR buffer reagents; thus, only template, PCR primers and PCR-grade water are added.
The Hot Start Taq Polymerase is inactive during reaction setup due to the bound antibody, which is quickly released at elevated temperatures, ensuring the enzyme is active only during PCR. There is no need for prolonged heating or denaturation steps. Thus, primer dimers and miss-priming do not occur. QUALITY CONTROL Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay
Hot Start PCR Master Mix: Functional PCR Assay
Red Hot Start DNA Polymerase, 5 U/µl
CAT. # CONTENTS
BR0200301 100 U
20 µl Red Hot Start DNA Polymerase, 5 U/µl 1.8 ml 10x Reaction Buffer
1.5 ml 50 mM MgCl2
1.8 ml 5x PCR Booster BR0200302 500 U
100 µl Red Hot Start DNA Polymerase, 5 U/µl 2 x 1.8 ml 10x Reaction Buffer
1.5 ml 50 mM MgCl2
2 x 1.8 ml 5x PCR Booster BR0200303 5 x 500 U
5 x 100 µl Red Hot Start DNA Polymerase, 5 U/µl
5 x 1.8 ml 10x Reaction Buffer 5 x 1.5 ml 50 mM MgCl2
5 x 1.8 ml 5x PCR Booster APPLICATIONS
High-throughput, simplified PCR setup with easy identification of reaction mixtures that contain enzyme High specificity, hot-start amplification up to 4 kb Amplification of low-copy-number targets TA cloning
FEATURES
Red Hot Start DNA Polymerase Storage Buffer allows mixtures containing the enzyme to be identified
Highest PCR specificity and sensitivity without prolonged reactivation
COMPOSITION OF COMPONENTS Red Enzyme Storage Buffer:
Proprietary composition, 5 U/µl Hot Start Taq DNA Polymerase in a red-colored Storage Buffer 10x Reaction Buffer:
670 mM Tris-HCl (pH 8.8 at 25°C), 166 mM (NH4)2SO4,
4.5% Triton® X-100, 2 mg/ml gelatin
DESCRIPTION
biotechrabbit Red Hot Start DNA Polymerase is a first -choice hot-start enzyme ensuring high product yields with low background and simplified PCR setup. This red-colored enzyme formulation performs in all respects like colorless Hot Start Taq DNA Polymerase and ensures low-background, high-specificity amplification without primer dimer formation and miss-priming. Additionally, reaction mixtures containing the enzyme are easily identified due to the red-colored enzyme Storage Buffer.
The Hot Start Taq Polymerase is inactive during reaction setup due to the bound antibody, which is quickly released at elevated temperatures, ensuring the enzyme is active only during PCR. There is no need for prolonged heating or denaturation steps.
Thus, primer dimer formation and miss-priming do not occur. The PCR specificity can be further increased in some cases by addition of the supplied 5x PCR Booster. The optional use of 5x PCR Booster improves PCR results in many cases, including impure template or low template amounts. Taq DNA polymerase is a thermostable, highly processive 5'→3' DNA polymerase that has low 5'→3' exonuclease activity and lacks 3'→5' exonuclease (proofreading) activity. The enzyme also exhibits deoxynucleotidyl transferase activity that results in the addition of an extra A overhang at the 3' ends of PCR products allowing easy cloning of PCR products into vectors with T overhangs.
UNIT DEFINITION
One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol of dNTP into acid -insoluble form in 30 minutes at 72°C in the presence of the reaction buffer. QUALITY CONTROL Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Functional PCR Assay
Red Hot Start PCR Master Mix, 2x
CAT. # CONTENTS
BR0200401 100 reactions (50 µl) 2 x 1.25 ml Red Hot Start PCR Master Mix, 2x BR0200402 500 reactions (50 µl) 10 x 1.25 ml Red Hot Start PCR Master Mix, 2x APPLICATIONS
High-throughput, simplified PCR setup with easy
identification of the reaction mixtures that contain enzyme Hot-start PCR amplification up to 4 kb
Amplification of low-copy-number targets TA cloning
FEATURES
Exceptionally pure Hot Start Taq DNA Polymerase and highest quality dNTPs in a red-colored master mix for simplified reaction setup
Highest PCR specificity and sensitivity without prolonged reactivation
COMPOSITION OF COMPONENTS Red Hot Start PCR Master Mix, 2x:
Proprietary composition, contains all necessary PCR reagents, including 3 mM MgCl2 (1x has 1.5 mM MgCl2) and
the red dye
DESCRIPTION
biotechrabbit Red Hot Start PCR Master Mix is a perfect choice for simplified hot-start PCR setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. Additionally the colored formulation allows the reactions that contain the enzyme and dNTPs to be identified.
This red-colored master mix performs in all respects like colorless Hot Start PCR Master Mix and ensures high product yields from various templates with low background.
The 2x Red Hot Start PCR Master Mix contains pure
biotechrabbit Hot Start Taq DNA polymerase, highest quality dNTPs and optimal colored PCR buffer reagents; thus, only template, PCR primers and PCR-grade water are added. The Hot Start Taq Polymerase is inactive during reaction setup due to the bound antibody, which is quickly released at elevated PCR temperatures, ensuring the enzyme is active only during PCR. There is no need for prolonged heating or denaturation steps. Thus, primer dimer formation and miss-priming do not occur.
QUALITY CONTROL Components:
Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay
Red Hot Start Master Mix: Functional PCR Assay
Green Hot Start PCR Master Mix, 2x
CAT. # CONTENTS
BR0200501 100 reactions (50 µl) 2 x 1.25 ml Green Hot Start PCR Master Mix, 2x BR0200502 500 reactions (50 µl) 10 x 1.25 ml Green Hot Start PCR Master Mix, 2x APPLICATIONS
High-specificity, high-throughput hot-start PCR for direct loading onto gels
Amplification of low-copy-number targets TA Cloning
FEATURES
Green Hot Start PCR Master Mix formulation allows direct loading onto gels after PCR
Highest PCR specificity and sensitivity without prolonged reactivation
Exceptionally pure Hot Start Taq DNA Polymerase and highest quality dNTPs in a 2x Hot Start PCR Master Mix
COMPOSITION OF COMPONENTS Green Hot Start PCR Master Mix,2x:
Proprietary composition contains all necessary PCR reagents, including 3 mM MgCl2 (1x has 1.5 mM MgCl2) and green and
yellow electrophoresis dyes
DESCRIPTION
biotechrabbit Green Hot Start PCR Master Mix is a perfect choice for a fast reaction setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. It is designed for low-background, high-throughput PCR amplification of 0.2 4 kb DNA targets. Additionally the special formulation allows reactions to be loaded directly onto gels after amplification without adding additional loading dye.
The 2x Hot Start PCR Master Mix contains pure biotechrabbit Hot Start Taq DNA polymerase, highest quality dNTPs and optimal Green PCR Buffer reagents; thus, only template, PCR primers and PCR-grade water are added. Green Reaction Buffer also allows mixtures containing the enzyme to be identified.
The Hot Start Taq Polymerase is inactive during reaction setup due to the bound antibody, which is quickly released at elevated temperatures, ensuring the enzyme is active only during PCR. There is no need for prolonged heating or denaturation steps. Thus, the primer dimer formation and miss-priming do not occur.
Green Hot Start PCR Master Mix contains two dyes (blue and yellow) that separate during electrophoresis, allowing migration progress to be monitored. Reactions with Green Hot Start PCR Master Mix have sufficient density for direct loading onto agarose gels. The dyes absorb between 225 300 nm, making standard A260 readings to determine DNA
concentration unreliable. QUALITY CONTROL Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay
Green Hot Start PCR Master Mix: Functional PCR Assay
High Fidelity and Long Range PCR Product Selection Guide
The PCR accuracy achievable with Taq DNA Polymerase (~one error per 45,000 nucleotides incorporated) is not always sufficient. For demanding cloning, sequencing and expression applications , polymerases w →
activity) are preferred. These proofreading enzymes can reduce PCR error rates by 10 fold or more, depending on the polymerase. Pfu DNA Polymerase, originally isolated from Pyrococcus furiosus, is one of the most commonly used proofreading enzymes. The unmodified form exhibits approximately 8–10 times higher accuracy than Taq DNA Polymerase. PCR fidelity, however, depends greatly on reaction conditions, including pH, magnesium concentration, balance of dNTPs, base composition of the template and purity of the components.
When working with Pfu DNA polymerase, be aware that its → might cause primer degradation. Generally, primers for use with the Pfu enzyme should be slightly longer than those for Taq, and the enzyme should always be the last reagent added to the reaction mixture. Pfu is slower than Taq and elongation of 1 kb requires 2 minutes, whereas Taq polymerase typically requires 1 minute.
The synergy of Taq DNA polymerase and a proofreading DNA polymerase is used to generate longer PCR products with higher accuracy than that of Taq alone. Such enzymes blends can be used for the amplification of templates of up to 30 40 kb in length and of GC-rich templates. In the latter case, different additives (such as DMSO, formamide or betaine) are helpful to enhance the denaturation of GC-rich templates for higher yields.
Product Main feature
Main application Fidelity Maximum product length GC-rich templates Simplified setup Optimization freedom Blunt Cloning Use of dUTP High yield Pfu DNA Polymerase, 2.5 U/µl
Ten times more accurate than Taq DNA Polymerase; supplied with PCR Helper for better results High-fidelity ~10x higher than Taq 3-5 kb
Not recommended Pfu PCR Master Mix, 2xIncludes Pfu DNA Polymerase and high-fidelity PCR reagents High-throughput, high-fidelity ~10x higher than Taq 3 kb
Not recommendedLong and High Fidelity PCR Enzyme Mix, 2.5 U/µl A blend of thermostable DNA Polymerases for long-range and high-fidelity PCR, with PCR Helper for better results with GC-rich templates Long-range, GC-rich templates Similar to
Pfu >30 kb
requires blunting recommended Not
Long Range PCR Master Mix, 2x Includes a blend of thermostable DNA Polymerases and reagents for long-range and high-fidelity PCR and GC-rich templates High-throughput, long-range, GC-rich
Pfu DNA Polymerase, 2.5 U/µl
CAT. # CONTENTS
BR0300101 100 U 40 µl Pfu DNA Polymerase, 2.5 U/µl 0.5 ml 10x Pfu Reaction Buffer 0.25 ml 5x PCR Helper
BR0300102 500 U 200 µl 2.4 ml 10x Pfu Reaction Buffer Pfu DNA Polymerase, 2.5 U/µl 1.2 ml 5x PCR Helper
APPLICATIONS High-fidelity PCR
Blunt cloning of PCR products Site-directed mutagenesis FEATURES
Accurate PCR for demanding applications
Approximately 10 times higher accuracy than that of Taq DNA Polymerase
COMPOSITION OF COMPONENTS
2.5 U/µl Pfu DNA Polymerase in Storage Buffer:
50 mM Tris-HCl (pH 8.0 at 25C), 1 mM DTT, 0.1 mM EDTA, 0.1% (v/v) Nonidet P-40, 1 mM PMSF, 0.1% (v/v) Tween 20, 50% (v/v) glycerol
10x Pfu Reaction Buffer:
500 mM Tris-HCl, 140 mM (NH4)2SO4, 17.5 mM MgCl2,
pH 9.1 at 25C.
DESCRIPTION
biotechrabbit Pfu DNA Polymerase is a highly purified recombinant thermostable proofreading DNA polymerase. Pfu DNA Polymerase exhibits approximately 10 times higher accuracy than Taq DNA Polymerase.
The enzyme catalyzes template-dependent nucleotide polymerization in the 5'→3' direction. Additionally it exhibits 3'→5' exonuclease (proofreading) activity that allows the correction of nucleotide incorporation errors, thereby increasing fidelity and accuracy of PCR.
The enzyme has no 5'→3' exonuclease activity and no detectable reverse transcriptase activity.
For the most demanding applications, the supplied 5x PCR Helper can be optionally used for improving results when using templates with GC-rich sequences and complex structures. The use of dUTP and dITP is not recommended with Pfu, because Pfu binding to DNA templates with uracil and hypoxanthine stalls DNA synthesis.
Pfu DNA Polymerase produces blunt-end PCR products for blunt cloning.
UNIT DEFINITION
One unit catalyzes the incorporation of 10 nmol of dNTPs into a polynucleotide fraction in 30 minutes at 72C.
QUALITY CONTROL
Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay
Functional PCR Assay
Pfu PCR Master Mix, 2x
CAT. # CONTENTS
BR0300201 100 reactions (50 µl) 2 x 1.25 ml Pfu PCR Master Mix, 2x 0.25 ml 5x PCR Helper BR0300202 500 reactions (50 µl) 10 x 1.25 ml 1.2 ml 5x PCR Helper Pfu PCR Master Mix, 2x APPLICATIONS
High-throughput, high-fidelity PCR Blunt cloning of PCR products FEATURES
Pure Pfu DNA Polymerase and highest quality dNTPs in a 2x Pfu PCR Master Mix formulation
Accurate PCR for demanding applications
Approximately 10 times higher accuracy than that of Taq DNA Polymerase
COMPOSITION OF COMPONENTS Pfu PCR Master Mix, 2x:
Proprietary composition contains all necessary PCR reagents
DESCRIPTION
biotechrabbit Pfu PCR Master Mix is a perfect choice for fast high-fidelity PCR setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. It is designed for routine throughput, high-fidelity amplification of targets up to 3-4 kb in size.
The 2x Pfu PCR Master Mix contains Pfu DNA polymerase, highest quality dNTPs and optimal PCR buffer reagents; thus, only template, PCR primers and PCR-grade water are added. For the most demanding applications, the supplied 5x PCR Helper can be optionally be used to improve results when using templates with GC-rich sequences and complex structures.
Pfu DNA Polymerase exhibits approximately 10 times higher accuracy compared to Taq DNA Polymerase.
The use of dUTP and dITP is not recommended with Pfu, because Pfu binding to DNA templates with uracil and hypoxanthine stalls DNA synthesis.
Pfu DNA Polymerase produces blunt-end PCR products suitable for blunt cloning.
QUALITY CONTROL Components:
Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay
Pfu PCR Master Mix: Functional PCR Assay
Long And High Fidelity PCR Enzyme Mix, 2.5 U/µl
CAT. # CONTENTS
BR0300301 100 U
40 µl Long and High Fidelity PCR Enzyme Mix, 2.5 U/µl
0.5 ml 10x HF Reaction Buffer 0.25 ml 5x PCR Helper BR0300302 500 U
200 µl Long and High Fidelity PCR Enzyme Mix, 2.5 U/µl
2.4 ml 10x HF Reaction Buffer 1.2 ml 5x PCR Helper APPLICATIONS
High-fidelity, long-range PCR Amplification of GC-rich templates FEATURES
Accurate, long-range amplification up to 30 kb Accurate amplification of GC-rich templates COMPOSITION OF COMPONENTS
2.5 U/µl thermophilic polymerase blend in storage buffer: 20 mM Tris-HCl (pH 8.0 at 25C), 100 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.1% (v/v) Nonidet P-40, 1 mM PMSF, 0.1% (v/v) Tween 20, 50% (v/v) glycerol 10x HF Reaction Buffer: 500 mM Tris-HCl, 140 mM (NH4)2SO4, 17.5 mM MgCl2, pH 9.1 at 25C. DESCRIPTION
biotechrabbit Long and High Fidelity PCR Enzyme Mix is a first-choice enzyme blend for amplification of targets up to 40 kb in size with higher accuracy than that of Taq DNA
Polymerase.
This specially designed blend of thermophilic polymerases is well suited for amplification of targets that are GC-rich and have complex structures.
For the most demanding applications, the supplied 5x PCR Helper can be optionally used to improve results when using templates with GC-rich sequences and complex structures. Long and High Fidelity PCR Enzyme Mix produces a mixture of A-tailed and blunt-end PCR products: it is advisable to blunt products before cloning into blunt-end vectors.
UNIT DEFINITION
One unit catalyzes the incorporation of 10 nmol of
deoxyribonucleotides into a polynucleotide fraction (adsorbed on DE-81) in 30 minutes at 72C. QUALITY CONTROL Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Functional PCR Assay
Long Range PCR Master Mix, 2x
CAT. # CONTENTS
BR0300401 100 reactions (50 µl) 2 x 1.25 ml Long Range PCR Master Mix, 2x 0.25 ml 5x PCR Helper
BR0300402 500 reactions (50 µl) 10 x 1.25 ml Long Range PCR Master Mix, 2x 1.2 ml 5x PCR Helper
APPLICATIONS
High-throughput, long-range PCR Accurate amplification of more than 30 kb Blunt cloning of PCR products
FEATURES
Long-range and accurate amplification of more than 30 kb A blend of thermophilic polymerases and highest quality
dNTPs in a convenient 2x master mix formulation COMPOSITION OF COMPONENTS
Long Range PCR Master Mix, 2x:
Proprietary composition contains a blend of thermophilic DNA polymerases and all necessary PCR reagents
DESCRIPTION
biotechrabbit Long Range PCR Master Mix is a perfect choice for fast reaction setup for long-range PCR that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. It is designed for amplification of targets up to 30 kb in size. The master mix works well with GC-rich templates and amplifies DNA with a higher fidelity than that of Taq DNA Polymerase.
The Long Range PCR Master Mix contains a blend of thermophilic polymerases, highest quality dNTPs and optimal PCR buffer reagents; thus, only template, PCR primers and PCR-grade water are added.
For the most demanding applications, the supplied 5x PCR Helper can be optionally used to improve results when using templates with GC-rich sequences and complex structures. Long Range PCR Master Mix produces a mixture of A-tailed and blunt-end PCR products: it is advisable to blunt products before cloning into the blunt-end vector.
QUALITY CONTROL Components:
Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay
Long Range PCR Master Mix: Functional PCR Assay
Reverse Transcription and RT-PCR Product Selection Guide
The basic difference between PCR and RT-PCR is the template: DNA is the PCR template and RNA is the RT-PCR template. The first step in RT-PCR is the synthesis of single-stranded antisense DNA from an RNA template. This process employs reverse
transcriptases (RNA-dependent DNA polymerases). The Moloney-Murine Leukemia Virus (M-MuLV) reverse transcriptase is a classic example of such an enzyme.
Mutations have been introduced into reverse transcriptase genes to increase performance. biotechrabbit offers a modified reverse transcriptase, RevertUP Reverse Transcriptase, which is a proprietary modification of the M-MuLV reverse transcriptase with point mutations in polymerase and RNAse H domains. These mutations ensure elevated cDNA synthesis efficiency and eliminate RNase H activity,
cDNA can be synthesized by reverse transcriptase either using random primers, oligo -dT primers or sequence-specific primers. Special care should be taken to prevent RNA templates from degradation by pervasive RNAses. Therefore it is always a safe approach to use RNAse inhibitors in all reactions involving RNA.
RT-PCR can be performed in one step or two steps. Two-step RT-PCR includes two different experiments, first the reverse transcription of RNA template is performed and then the PCR amplification of the cDNA template is performed in another tube. One -step RT-PCR combines both reactions in one tube: cDNA is synthesized from RNA and then amplified to generate cDNA with specific primers without additional pipetting steps.
Taq DNA Polymerase and its hot-start versions are the main enzymes used for cDNA amplification in both the one-step and two-step RT-PCR.
Product Application Main feature
One Step
RT-PCR Kit One-step RT-PCR
A blend of efficient thermostable reverse transcriptase and proprietary Ribonuclease Inhibitor provides high cDNA yields.
Unique Hot Start Taq DNA Polymerase in a mix with high-quality dNTPs and PCR enhancers allows sensitive, low-background amplification.
RevertUP Reverse Transcriptase Reliable synthesis of , two-step RT-PCR
A proprietary M-MuLV reverse transcriptase engineered by point mutations in polymerase and RNAse H domains that ensure efficient cDNA synthesis with no RNAse H activity, allowing succe
MMuLV Reverse Transcriptase
cDNA synthesis,
two-step RT-PCR A classical, exceptionally pure reverse transcriptase supplied with Reaction Buffer.
RNAse Inhibitor
Prevention of RNA degradation by RNAses
A recombinant, non-competitive inhibitor of pancreatic-type ribonucleases such as RNAse A, RNAse B, and RNAse C.
One Step RT-PCR Kit
CAT. # CONTENTS
BR0400101 50 reactions (50 µl) 1250 µl One Step Mix, 2x 125 µl RT–RI Blend, 20x BR0400102 100 reactions (50 µl) 2 x 1250 µl One Step Mix, 2x 2 x 125 µl RT–RI Blend, 20x BR0400103 500 reactions (50 µl) 10 x 1250 µl One Step Mix, 2x 10 x 125 µl RT–RI Blend, 20x APPLICATIONS One-step RT-PCR Cloning Expression-level determination Virus detection FEATURES
A blend of efficient reverse transcriptase and proprietary Ribonuclease Inhibitor provides high cDNA yields Unique Hot Start Taq DNA Polymerase in a mix with
high-quality dNTPs and PCR enhancers allows sensitive, low-background amplification
Successful amplification of GC-rich and complex templates
COMPOSITION OF COMPONENTS One Step Mix, 2x:
Proprietary composition, a mix of Hot Start Taq Polymerase, 6 mM MgCl2, 2 mM dNTPs, enhancers and
stabilizers
RT-RI Blend, 20x:
Proprietary composition, a blend of efficient reverse transcriptase and unique Ribonuclease Inhibitor
DESCRIPTION
biotechrabbit One Step RT-PCR Kit provides an easy and efficient way to perform both reverse transcription of RNA and cDNA amplification by PCR in one step.
Only RNA template, primers and PCR-grade water are added. The 20x RT RI Blend — a blend of an efficient thermostable reverse transcriptase and a proprietary Ribonuclease Inhibitor
— ensures high yields of cDNA.
The 2x One Step Mix, which contains unique Hot Start Taq DNA Polymerase, dNTPs, MgCl2 and stabilizers in an optimized
buffer, provides high PCR product yields with minimal backgrounds even from low-abundance template and difficult targets.
The PCR enhancers included in the mix allow efficient amplification of complex templates including GC - or AT-rich sequences.
QUALITY CONTROL Functional RT-PCR Assay
NEW
RevertUP
™
Reverse Transcriptase, 100 U/µl
CAT. # CONTENTS BR0400301 10000 U for 100 reactions (20 µl) 100 µl RevertUP™ Reverse Transcriptase, 100 U/µl 1 ml 5x RevertUP™ Buffer BR0400302 50000 U for 500 reactions (20 µl) 500 µl RevertUP™ Reverse Transcriptase, 100 U/µl 5 x 1 ml 5x RevertUP™ Buffer APPLICATIONS
First-strand cDNA synthesis cDNA labeling
RNA analysis by primer extension FEATURES
Efficient at high temperatures (up to 55oC)
High-sensitivity cDNA synthesis from a few template copies
Exceptionally pure enzyme for demanding applications COMPOSITION OF COMPONENTS
RevertUP Reverse Transcriptase 100 U/µl in Storage Buffer: 50 mM Tris-HCl (pH 7.5 at 25°C), 100 mM NaCl, 0.1 mM EDTA, 10 mM DTT, 0.01% Nonidet P-40, 50% glycerol 5x RevertUP Buffer: Proprietary, contains 25 mM Mg2+ DESCRIPTION
RevertUP is a proprietary MMuLV reverse transcriptase engineered by point mutations in polymerase and RNase H domains. This ensures efficient cDNA synthesis and eliminate RNase H activity,
cDNA.
Reverse transcriptase is a DNA polymerase which utilizes RNA as a substrate and exhibits no measurable proofreading 3 '→5' exonuclease function. Enzyme performs cDNA synthesis by extending a DNA primer annealed to an RNA template or copies ssDNA template.
UNIT DEFINITION
One unit is the amount of enzyme activity that incorporates 1 nmole of dTTP into acid insoluble fraction in 10 minutes at 42°C when poly A+ RNA and oligo-dT(20) are used as
template primer. QUALITY CONTROL
RNAse Assay Functional Assay
RevertUP™ RTase X RTase Y
Picture 1. RT-PCR of the G3PDH gene (500 bp). cDNA templates were synthesized
with various RNAse H reverse transcriptases from G3PDH mRNA (102 105
copies/reaction). The reverse transcription reaction was performed with specific reverse primers and 100 U enzyme at 42ºC for 20 minutes
MMuLV Reverse Transcriptase, 200 U/µl
CAT. # CONTENTS
BR0400201 10000 U for 50 reactions (10 µl)
50 µl MMuLV Reverse Transcriptase, 200 U/µl
1 ml 10x MMuLV RT Buffer BR0400202 50000 U for 250 reactions
(10 µl)
250 µl MMuLV Reverse Transcriptase, 200 U/µl
1 ml 10x MMuLV RT Buffer APPLICATIONS
First-strand cDNA synthesis Generation of labeled cDNA RNA analysis by primer extension FEATURES
High yields of first-strand cDNA
Exceptionally pure reverse transcriptase for demanding applications
COMPOSITION OF COMPONENTS
MMuLV Reverse Transcriptase, 200 U/µl in Storage Buffer: 50 mM Tris-HCl, 150 mM NaCl, 0.1 mM EDTA,1 mM DTT, 0.1% NP-40 Alternative, 50% glycerol, pH 7.6 at 25°C 10x MMuLV RT Buffer:
500 mM Tris-HCl, 750 mM KCl, 30 mM MgCl2, 100 mM
DTT, pH 8.3 at 25°C
DESCRIPTION
biotechrabbit MMuLV Reverse Transcriptase is an
exceptionally pure DNA polymerase which utilizes RNA as a substrate and exhibits no measurable proofreading 3 '→5' exonuclease function. This enzyme can perform cDNA synthesis by extending a DNA primer annealed to an RNA template, or can copy a single-stranded DNA template. The enzyme is purified from a recombinant E. coli strain carrying the MMuLV reverse transcriptase gene. UNIT DEFINITION
One unit is defined as the amount of enzyme required to incorporate 1 nmol of dTTP into acid insoluble material in 10 minutes at 37°C using poly rA/oligo-dT as a substrate. QUALITY CONTROL
Single-Stranded Exonuclease Activity Double-Stranded Exonuclease Activity Double-Stranded Endonuclease Activity E. coli 16S rDNA Contamination Test Non-Specific RNAse Assay
RNAse Inhibitor, 40 U/µl
CAT. # CONTENTS
BR0400901 2500 U 62.5 µl RNAse Inhibitor, 40 U/µl BR0400902 10000 U 250 µl RNAse Inhibitor, 40 U/µl APPLICATIONS
Prevention of RNA degradation by Ribonucleases A, B or C during:
In vitro transcription and translation cDNA synthesis
RNA purification and storage FEATURES
Exceptionally pure proprietary Ribonuclease Inhibitor for demanding RNA applications
Active under variety of reaction conditions for RNA COMPOSITION OF COMPONENTS
RNAse Inhibitor, 40 U/µl in Storage Buffer:
20 mM Hepes-KOH, 50 mM KCl, 8 mM DTT, 50% glycerol, pH 7.5 at 25°C
DESCRIPTION
biotechrabbit RNAse Inhibitor is an acidic, 52 kDa protein that is a potent non-competitive inhibitor of pancreatic-type ribonucleases such as RNase A, RNase B, and RNase C. The enzyme is provided as a fusion of the porcine RNAse Inhibitor gene with a proprietary, 22.5 kDa protein tag.
The protein is purified from a recombinant E. coli strain carrying the porcine RNAse Inhibitor gene.
UNIT DEFINITION
One unit is defined as the amount of enzyme required for 50%
inhibition of -cyclic
monophosphate by 5 ng of RNAse A. QUALITY CONTROL
Single-Stranded Exonuclease Activity Double-Stranded Exonuclease Activity Double-Stranded Endonuclease Activity E. coli 16S rDNA Contamination Test Non-Specific RNAse Assay
Quantitative Real-Time PCR Product Selection Guide
The difference between quantitative real-time PCR (qPCR) and end-point PCR is the step at which the PCR product amount is measured. In traditional PCR, product quantitation is performed once at the end of the reaction. In qPCR, the amount of PCR product is measured after every amplification round. qPCR instruments are designed for both PCR cycling and the detection of signal from fluorescent dyes during amplification. By monitoring the progress of amplification in real time , the point in time when the
amplification of the target is detected above background can be identified. The fewer targets present in the reaction, the later the first significant signal is detected. This technique , therefore, is of great advantage when gene-expression differences are analyzed. There are two ways to detect PCR signal with the help of fluorescent dyes: fluorescent dyes that nonspecifically bind to double -stranded DNA or fluorescently labeled, sequence-specific probes.
The use of universal fluorescent dyes enables analysis of many targets without having to synthesize sequence -specific probes. However, this method has the disadvantage of detecting all specific and unspecific amplification events as well as primer dimers because the dye binds to all double-stranded DNA. To achieving high PCR specificity, avoiding background and primer dimer formation is critical. The fluorescent-probe method is more precise, as the use of sequence -specific fluorescently labeled probes ensures the detection of the target product only. A variety of probe fluorescence chemistry is currently available. Multiple fluorescent probes can be used in multiplex qPCR, although the use of specific probes is more expensive than using DNA-binding dyes.
QPCR OneStep Kits allow reverse transcription and real-time PCR to be performed in one step. In this case, RNA is the template for qPCR. Some one-step kits for qPCR provide an intercalating dye for probe detection. One-step qPCR kits include separate qPCR and reverse transcription master mixes.
Many real-time PCR instruments available on the market require a passive internal reference dye (either ROX™ or fluorescein) for normalization of the fluorescent signal. The use of passive reference dyes does not interfere with the qPCR.
The table below will help you select a qPCR master mix for use on your instrument:
Real-time PCR master mixes (for fast or standard mode and for multiplex PCR)
Passive
reference dye Instruments Intercalating-dye-based mixes (SYBR®
Green I, EvaGreen or other), such as QPCR Green Master Mix
Nonspecific, detects all double-stranded DNA
Cheaper and faster
ROX Applied Biosystems: ABI PRISM
® 7000, 7300, 7900 HT, and StepOne for
7500, ROX solution should be provided separately Fluorescein Bio-Rad® iCycler® iQ and iQ 5
None
Roche LightCycler® 480, Roche LightCycler® 1.5, 2.0, Bio-Rad® ®
SmartCycler®, QIAGEN® Rotor-Gene® 3000, Rotor-Gene® 6000, Eppendorf®
Mastercycler® ep REALPLEX® and Techne® Quantica®
Probe-based mixes (TaqMan®, Molecular
Beacon® or other), such as QPCR Probe Master Mix
Can be used specifically Can be universally More expensive
Takes more time to design and synthesize specific probes
ROX Applied Biosystems: ABI PRISM
® 7000, 7300, 7900 HT, and ; for
7500, ROX solution should be provided separately
Fluorescein Bio-RadRad® CFX96 or Roche LightCycler® iCycler® iQ ®5, QIAGEN 480. ® Rotor-Gene® 6000,
Bio-None
Roche LightCycler® 480, Roche LightCycler® 1.5, 2.0, Bio-Rad® ®
SmartCycler®, QIAGEN® Rotor-Gene® 3000, Rotor-Gene® 6000, Eppendorf®
biotechrabbit qPCR master mixes that do not contain the reference dye in the mix (reference dye can be supplied separately): Product
Reference
dye supplied Application Instruments
QPCR Green Master Mix, 2x
None Intercalating-dye-based qPCR
Roche LightCycler® 480, Roche LightCycler® 1.5, 2.0, Bio-Rad® ®
SmartCycler®, QIAGEN® Rotor-Gene® 3000, Rotor-Gene® 6000, Eppendorf®
Mastercycler® ep REALPLEX® and Techne® Quantica®
ROX ,
separate Intercalating-dye-based qPCR on ROX instruments Applied Biosystems: ABI PRISM
® 7000, 7300, 7900 HT and ; for
7500, ROX solution should be provided separately Fluorescein,
separate Intercalating-dye-based qPCR on fluorescein instruments Bio-Rad® iCycler® iQ 5 QPCR
Probe Master Mix, 2x
None Probe-based qPCR
Roche LightCycler® 480, Roche LightCycler® 1.5, 2.0, Bio-Rad® ®
SmartCycler®, QIAGEN® Rotor-Gene® 3000, Rotor-Gene® 6000, Eppendorf®
Mastercycler® ep REALPLEX® and Techne® Quantica®
ROX ,
separate Probe-based qPCR on ROX instruments Applied Biosystems: ABI PRISM
® 7000, 7300, 7900 HT and StepOne for
7500, ROX solution should be provided separately Fluorescein,
separate Probe-based qPCR on fluorescein instruments Bio-Rad
® iCycler® iQ 5, QIAGEN® Rotor-Gene® 6000,
Bio-Rad® CFX96 and Roche LightCycler® 480
biotechrabbit qPCR master mixes that contain ROX™ (low or high concentration) in the master mix:
Product Reference dye supplied Company Instrument
QPCR SyGreen Master Mix, LRox and QPCR Probe-R Master Mix, LRox Low-concentration ROX , in mix Analytica Jena Applied Biosystems Bio-Rad Cepheid Eppendorf Illumina QIAGEN/Corbett Roche Applied Science Stratagene (Agilent) Takara
Techne
qTower
7500, 7500 FAST, Viia 7
iCycler®, MyiQ , iQ 5, Opticon , Opticon 2, Chromo4 ,
MiniOpticon , CFX96 , CFX384 Smartcycler®
Mastercycler® ep REALPLEX®, Mastercycler® REALPLEX® 2S
Eco
Rotor-Gene® 3000, 6000, Q
Lightcycler®480, Lightcycler® Nano
Mx4000P®, Mx3000P®, Mx3005P® Cycler Dice® Quantica® QPCR SyGreen Master Mix, HRox and QPCR Probe-R Master Mix, HRox High concentration ROX , in mix Analytica Jena Applied Biosystems Cepheid Eppendorf Illumina QIAGEN/Corbett Roche Applied Science Takara Techne qTower 7000, 7300,7700,7900, 7900HT, 7900HT FAST, StepOne , StepOne Plus Smartcycler®
Mastercycler® ep REALPLEX®, Mastercycler® REALPLEX® 2S
Eco
Rotor-Gene® 3000, 6000, Q
Lightcycler®480, Lightcycler® Nano
Cycler Dice®
Quantica®
biotechrabbit QRT-PCR OneStep Kits: QRT-PCR SyGreen
or
Probe-R One Step Kits, LROX
QRT-PCR SyGreen or
Probe-R One Step Kits,
HROX Company Instrument
Yes Yes Analytica Jena qTower
Yes NO Applied Biosystems 7500, 7500 FAST, Viia 7
NO Yes Applied Biosystems 7000, 7300,7700,7900, 7900HT, 7900HT FAST, StepOne , StepOne Plus Yes NO Bio-Rad iCyclerMiniOpticon , CFX96 , CFX384 ®, MyiQ , iQ 5, Opticon , Opticon 2, Chromo4 ,
Yes Yes Cepheid Smartcycler®
Yes Yes Eppendorf Mastercycler® ep REALPLEX®, Mastercycler® REALPLEX® 2S
Yes Yes Illumina Eco
Yes Yes QIAGEN/Corbett Rotor-Gene® 3000, 6000, Q
Yes Yes Roche Applied Science Lightcycler®480, Lightcycler® Nano
Yes NO Stratagene (Agilent) Mx4000P®, Mx3000P®, Mx3005P®
Yes Yes Takara Cycler Dice®
