Gradient Summary
- favorable conditions (temperature and buffer) is indicated by green highlighting, and corresponds to the green happy faces on each gel
- right-most column indicates number of primer pairs that work well under that condition
- will next try to multiplex 7 primer pairs using Buffer E at 64°C
- Scun2
- Scun3
- Scun9
- Scun10
- Scun11
- Scun15
- Scun22
Multiplex 1
- The first attempt at multiplexing!
PCR
- reaction conditions are based on the gradient summary (above) - Buffer E, 64°C
- run uniplexes at this reaction condition to check against the gradient PCR result, 4 replicates
- run multiplex at this reaction condition, 8 replicates
- hopefully, we'll get clear bands in both uniplexes and multiplex!
Gel
- Expected banding (based on gradient PCRs)
- Conclusion: missing the Scun22 band, but the other 6 look like they might be there
- ready to order labelled primers, and start multiplexing and fragging!
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