Multiplex 1
- SCOC multiplex diversity panel with labelled primers to send to frag! Woot!
PCR
Gel
- used left-over gel from 13 June 2011
Load |
Run
|
2 µL gel load dye + 4 µL PCR product |
160 V
|
6 µL ladder |
1 hour
|
- see 13 June 2011 for banding pattern expectations
- a whole lotta nada :(
- very strong primer band indicates no amplification took place
- might be a combination of:
- having to make primer dilutions last-second
- pipetting 0.8µL DNA
- using the flat-top-no-skirt PCR plate that didn't fit into the thermal cycler (and thus doing full-reaction transfer to cycler-friendly PCR plate)
- therefore taking a long time for PCR set-up
Next Step
- re-do PCR, but have 8-well strip with 0.8*4 µL DNA plus Big MM, then transfer from Mini MM straight across into PCR plate
- might also be worth doing a PCR similar to that from 13 June 2011 - uniplex of each primer pair, plus multiplex - to 'test' labeled primers
- over night, test full-skirt PCR plate with flat-top 8-well strips to see how much evaporation occurs - this is how I plan to ship for frag
- shit, now no matter how soon I can ship, it won't get to UAGC till Monday
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