Thursday, Nov. 13, 2014

In order to determine if ApeKI restriction enzyme will work for O. lurida, did test digestion with 4 individuals from different populations. Used ApeKI from Winston.

Test digestion with ApeKI

  • Determine concentration with Qubit (did not update standard from 11/5))
    • OR1_2: 4.78 ng/ul
    • W13_1: 6.82
    • BC4_17: 9.62
    • CA1_17: 96.4
  • Dry down in CentriVac at 60, ul for 200 ng:
    • B: 20.79
    • C: 2.07
    • O: 41.84
    • W: 29.32
  • ApeKI digestion at 75deg
    • 17 ul dH2O, 2 ul NEB3, 1ul ApeKI
    • 2:10p until 4:15pm
Sequencing Reaction of Sap/Exo
  • 86 reactions
    • well 16 from SapExo was empty (did not put lid on tight enough?)
      • BC3_11
  • Used cyc-seq program from Barber
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