The following pages illustrate how to develop a script for a typical Gemini application.
The chosen exercise is a column overlapping pooling application involving single pipetting using only standard tips. The purpose of the script is to pool two columns of the source plate into one column of the destination plate using single pipetting. The initial volume in the source plate is 200 µl and the pipetting volume is 50 µl (final total of 100 µl) per well in the destination plate.
You must first specify the worktable which is required. One microplate serves as the source plate and another microplate serves as the destination plate. Both plates are placed in the landscape orientation on the same MTP (microtiter plate or microplate) carrier. The application uses one wash station with shallow cleaner and waste.
Figure 9-1: Example script, step 1
For all normal applications using standard tips, it is necessary to start with a wash command. Click the Wash Tips button in the commands toolbar.
On the Wash dialog box that appears, select tips 1-8 (or click All Tips) and fill in the remaining fields as appropriate (appropriate values have already been chosen).
Click OK to add the properly defined Wash Tips command to your script.
Now click the Aspirate button in the commands toolbar. On the Aspirate dialog box that appears, select tips 1-8 (or click All Tips), specify 50 µl as the volume for all tips and click the source rack in the Worktable window. The source rack should then be highlighted in color. Now click on the first column in the Aspirate dialog box. Tips 1-8
Note:
Alternatively, press the Insert key on the keyboard and choose Wash Tips or choose Insert new Command in the Edit menu and choose Wash Tips.
Figure 9-2: Example script, step 2
should then be highlighted in green. Click them to select them and they will become green. The dialog box should now appear as follows:
Figure 9-3: Example script, step 3
Note:
You can customize the tip selection and tip spacing in the Aspirate and Dispense dialog boxes by clicking on the tips you want to use and choosing the spacing as required. For example, a tip spacing of 2 will select every other well.
Click OK to add the properly defined Aspirate command to your script. The program window should now appear as follows:
Now click the Dispense button in the commands toolbar. On the Dispense dialog box that appears, select tips 1-8 (or click All Tips), specify 50 µl as the volume for all tips and click the destination rack in the Worktable window. The destination rack should then be highlighted in color. Now click on the first column in the Dispense dialog box. Tips 1-8 Figure 9-4: Example script, step 4
should then be highlighted in green. Click them to select them and they will become green. The dialog box should now appear as follows:
Figure 9-5: Example script, step 5
Click OK to add the properly defined Dispense command to your script. The program window should now appear as follows:
• Now click the Wash Tips button in the commands toolbar. On the Wash dialog box that appears, select tips 1-8 (or click All Tips) and fill in the remaining fields as Figure 9-6: Example script, step 6
appropriate (suitable values are filled in automatically). The dialog box should now appear as follows:
Figure 9-7: Example script, step 7
Click OK to add the properly defined Wash Tips command to your script. The program window should now appear as follows:
Figure 9-8: Example script, step 8
• Now click the Begin Loop button in the commands toolbar. On the Begin Loop dialog box that appears, specify the number of loop cycles as 11 and give the loop the name Cycle 1.
Figure 9-9: Example script, step 9
Click OK to add the Begin Loop command to your script. The program window should now appear as follows:
• You must now define which actions have to be repeated 11 times. Because an Aspirate step should be executed first, click the Aspirate button in the commands toolbar and select tips 1-8 (or click All Tips), specify a pipetting volume of 50 µl, select the source rack and specify rack column 1 for the aspirate position.
Figure 9-10: Example script, step 1
• Additional loop options must now be defined. In this example, we will increase the rack column by +1 for every cycle in the loop.
Figure 9-11: Example script, step 11, showing loop options
Click OK to add the properly defined loop options for the Aspirate command to your script. The program window should now appear as follows:
• The next action to be executed inside the Loop Cycle 1 is a Dispense command.
Click the Dispense button in the commands toolbar, select tips 1-8 (or click All Tips), specify a pipetting volume of 50 µl, select the destination rack and specify rack Figure 9-12: Example script, step 12
column 2 for the dispense position. Additional loop options must now be defined. In this example, we will increase the rack column by +1 for every cycle in the loop.
Figure 9-13: Example script, step 13
Click OK to add the properly defined loop options for the Dispense command to your script. The program window should now appear as follows:
• The next action to be executed inside the Loop Cycle 1 is another Wash Tips command. Click the Wash Tips button in the commands toolbar, select tips 1-8 (or click All Tips), and fill in the remaining boxes in the Wash dialog box as appropriate (suitable values are filled in automatically). After the Wash Tips command has been Figure 9-14: Example script, step 14
fully specified (see the examples above in steps 1 and 4), click OK to add the command to your script. The program window should now appear as follows:
• The Wash Tips command is the last command you need in the loop Cycle 1. Click the End Loop button in the commands toolbar to add the End Loop command to the Figure 9-15: Example script, step 15
script. The End Loop command automatically refers to the previous Begin Loop command in the script. The program window should now appear as follows:
The Begin Loop Cycle 1 command (step 5) instructs the instrument to begin 11 cycles of the first loop of the script, which consists of steps 6-8 in the script. This instructs the instrument to aspirate from column 1 of the source rack, dispense to column 2 of the destination rack, aspirate from column 2 of the source rack, dispense to column 3 of the destination rack, and so on, until it aspirates from column 11 of the source rack and dispenses to column 12 of the destination rack.
• Now click the Aspirate button in the commands toolbar. On the Aspirate dialog box that appears, select tips 1-8 (or click All Tips), specify an Aspirate volume of 50 µl, Figure 9-16: Example script, step 16
select the source rack, and the position on the rack (column 12). The dialog box should now appear as follows:
Figure 9-17: Example script, step 17
Click OK to add the properly defined Aspirate command to your script. The program window should now appear as follows:
• Now click the Dispense button in the commands toolbar. On the Dispense dialog box that appears, select tips 1-8 (or click All Tips), specify a Dispense volume of 50 µl, Figure 9-18: Example script, step 18
select the destination rack, and the position on the rack (column 1). The dialog box should now appear as follows:
Figure 9-19: Example script, step 19
Click OK to add the properly defined Dispense command to your script. The program window should now appear as follows:
• The next step is to add another Wash Tips command. Since this Wash Tips
command is exactly the same as the Wash Tips commands defined previously, you can select (click on) the previously defined Wash Tips command in the script (it will be highlighted in color) and copy it using the Copy function in the Edit menu (or click the Copy button in the File toolbar). Then click the last line of the script (Dispense command, line 11) and choose Paste in the Edit menu (or click the Paste button in Figure 9-20: Example script, step 20
the File toolbar). This adds another Wash Tips command to the end of the script.
The program window should now appear as follows:
• Now click the Begin Loop button in the commands toolbar to add a new loop. On the Begin Loop dialog box that appears, specify the number of loop cycles as 11 and Figure 9-21: Example script, step 21
give the loop the nameCycle 2. Click OK to add the Begin Loop command to your script. The program window should now appear as follows:
• You must now define which actions have to be repeated 11 times. Because an Aspirate step should be executed first, click the Aspirate button in the commands toolbar and select tips 1-8 (or click All Tips), specify a pipetting volume of 50 µl, select the source rack and specify rack column 2 for the aspirate position. Additional Figure 9-22: Example script, step 22
loop options must now be defined. In this example, we will increase the rack column by +1 for every cycle in the loop. The dialog box should now appear as follows:
Figure 9-23: Example script, step 23
Click OK to add the properly defined loop options for the Aspirate command to your script:
• The next action to be executed inside the loopCycle 2is a Dispense command.
Click the Dispense button in the commands toolbar, select tips 1-8 (or click All Tips), specify a pipetting volume of 50 µl, select the destination rack and specify rack column 2 for the dispense position. Additional loop options must now be defined. In Figure 9-24: Example script, step 24
this example, we will increase the rack column by +1 for every cycle in the loop. The dialog box should now appear as follows:
Figure 9-25: Example script, step 25
Click OK to add the properly defined loop options for the Dispense command to your script. The program window should now appear as follows:
• The next item to add to the script is another Wash Tips command. To do this, click the Wash Tips button in the commands toolbar or copy a previous Wash Tips Figure 9-26: Example script, step 26
command to this line of the script as described above. The program window should now appear as follows:
• The Wash Tips command is the last command you need in the loop Cycle 2. Click the End Loop button in the commands toolbar to add the End Loop command to the Figure 9-27: Example script, step 27
script. The End Loop command automatically refers to the previous Begin Loop command in the script. The program window should now appear as follows:
The Begin Loop Cycle 2 command (step 13) instructs the instrument to begin 11 cycles of the second loop of the script, which consists of steps 14-16 in the script. This instructs the instrument to aspirate from column 2 of the source rack, dispense to column 2 of the destination rack, aspirate from column 3 of the source rack, dispense to column 3 of the destination rack, and so on, until it aspirates from column 12 of the source rack and dispenses to column 12 of the destination rack.
Figure 9-28: Example script, step 28
10 Validating, Starting and Stopping a Script
This chapter describes how to validate a script, locate errors in a script and start, pause, continue and stop a script.