User:Tara K. Luckau/Notebook/Team ConGen/2011/01/14

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Contents


Gel Lance conditions

Success!!!



  • Scun2 Primer - expecting bands ~150bp
    • Scun sample - band at 130bp
    • Scoc sample - band at 75bp


  • Scun22 Primer - expecting bands at ~450bp
    • Scun sample - multiple bands; but there is one prominent band in one of the replicates at 450bp
    • Scoc sample - multiple bands; but prominent band in all three replicates at 450bp


Differences between Lance conditions and previous trials

10January2011 (fail) 13January2011 (worked)
Buffer / MgCl2 1.5 µL 1 µL
Primers 0.1 µL each 0.25 µL each
Taq 0.05 µL (0.25 Units) 0.5 µL (2.5 Units)

The largest difference between the two reactions was Taq - 10-fold difference

  • In the interest of cost, try to pull this down to 1 Unit/reaction (0.2µL)
  • Also re-calculate for 10µL reaction volume
  • Find annealing temperatures (touchdowns are poopy)


PCR decrease Taq
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