BioMicroCenter:Wafergen: Difference between revisions
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== REACTION SETUP == | == REACTION SETUP == | ||
Wafergen reactions come in two flavors: prefilled arrays and "MyDesign". Prefilled arrays have specific primer sets laid out across the nanowells. MyDesign arrays are filled in two steps: cDNA followed by primers. <BR><BR> | Wafergen reactions come in two flavors: prefilled arrays and "MyDesign". Prefilled arrays have specific primer sets laid out across the nanowells. MyDesign arrays are filled in two steps: cDNA followed by primers. <BR><BR> | ||
Primers and cDNA are filled from specific 384 well plates in master mix already diluted to 1x. Technical replicates must be pipetted the desired number of times on the plate. eg. if you want to run your primers 3x on the cDNA, you should use 3 wells for each primer set and only 1 for each cDNA. Even though each dispense uses only 50nL, more material is required for pipetting. The matrix below outlines the preprogrammed sample/primer set ratios and the volumes required. | Primers and cDNA are filled from specific 384 well plates in master mix already diluted to 1x. Technical replicates must be pipetted the desired number of times on the plate. eg. if you want to run your primers 3x on the cDNA, you should use 3 wells for each primer set and only 1 for each cDNA. Even though each dispense uses only 50nL, more material is required for pipetting. The matrix below outlines the preprogrammed sample/primer set ratios and the volumes required. | ||
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|384 ||12ul ||12 ||18ul x4 wells | |384 ||12ul ||12 ||18ul x4 wells | ||
|} | |} | ||
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Wafergen recommends samples normalized to a concentration of 10 ng/uL. Although normalization is not required, the minimum acceptable concentration is 1 ng/uL.<br> | |||
Primers should be | Primers should be reconstituted to a concentration of 1.5 uM. | ||
== RUNNING THE CHIPS == | == RUNNING THE CHIPS == | ||
Revision as of 13:30, 7 March 2013
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The Wafergen SmartChip System is a highly flexible nanoliter qPCR instrument. The system is based on aluminum chips that contain a grid of 72x72 nanowells. Each well can hold up to 150ul of volume. The system itself is composed of two halves: a robot for loading the chips and a PCR machine for cycling and scanning the chips.
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REACTION SETUP
Wafergen reactions come in two flavors: prefilled arrays and "MyDesign". Prefilled arrays have specific primer sets laid out across the nanowells. MyDesign arrays are filled in two steps: cDNA followed by primers.
Primers and cDNA are filled from specific 384 well plates in master mix already diluted to 1x. Technical replicates must be pipetted the desired number of times on the plate. eg. if you want to run your primers 3x on the cDNA, you should use 3 wells for each primer set and only 1 for each cDNA. Even though each dispense uses only 50nL, more material is required for pipetting. The matrix below outlines the preprogrammed sample/primer set ratios and the volumes required.
| Samples | Primer Sets | ||
|---|---|---|---|
| Number | Volume | Number | Volume |
| 12 | 18ul x4 wells | 384 | 12ul |
| 24 | 18ul x2 wells | 216 | 12.5ul |
| 36 | 20.5ul | 144 | 13.5ul |
| 48 | 18ul | 108 | 14ul |
| 54 | 17.5ul | 96 | 14.5ul |
| 64 | 16.5ul | 80 | 15.5ul |
| 72 | 16ul | 72 | 16ul |
| 80 | 15.5ul | 64 | 16.5ul |
| 96 | 14.5 | 54 | 17.5ul |
| 108 | 14ul | 48 | 18ul |
| 144 | 13.5ul | 36 | 20.5ul |
| 216 | 12.5ul | 24 | 18ul x2 wells |
| 384 | 12ul | 12 | 18ul x4 wells |
Wafergen recommends samples normalized to a concentration of 10 ng/uL. Although normalization is not required, the minimum acceptable concentration is 1 ng/uL.
Primers should be reconstituted to a concentration of 1.5 uM.
RUNNING THE CHIPS
The Wafergen system is offered as a walkup service in the BioMicro Center. However, due to the complexity of the Wafergen system, a single brief training session is not sufficient for us to be comfortable letting users operate on their own. Instead, new users must use the robot under the guidance of the BioMicro Center technician for the first 2-3 runs. Once the user has completed their guided usage, you are free to use the system without supervision.
Users may sign up for the Wafergen system on the calendar. New users should confirm their schedule with BioMicro Center staff (biomicro@mit.edu).
DATA ANALYSIS
