User:Tara K. Luckau/Notebook/Team ConGen/2011/06/22: Difference between revisions

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* see 13 June 2011 for banding pattern expectations
* see 13 June 2011 for banding pattern expectations
* [[Image:20110622 Gel.jpg|600 px]]
* 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 transfer to 8-well strip with 0.8*4 µL DNA, then transfer 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




Revision as of 15:06, 22 June 2011

Tara's Lab Notebook <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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Multiplex 1

  • SCOC multiplex diversity panel with labelled primers to send to frag! Woot!

PCR


Gel

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 transfer to 8-well strip with 0.8*4 µL DNA, then transfer 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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