Tuesday 11/24/15

Finished gel extraction of samples B3-C4 from 2bRAD Library 1 started on 11/23/15 following the protocol described in that post.

Did the PCR of the rest of Library 1 (B5-C12) with the new Taq. There is no H9, however (due to how I labelled on the spreadsheet).

First made more Lib 1 and Lib 2 primer stock:

  • 10 uL stock + 90 uL NFW = 10 uM

PCR master mix. Added 3.75 of each 1 uM barcode (HT and BC) to wells.

1x
65
10 mM (each) dNTPS
1.5
97.5
10 uM ILL-Lib1
1.5
97.5
10 uM ILL-Lib2
1.5
97.5
5X Q5 buffer
15
975
Q5 Taq polymerase
.75
48.75
20.25 + barcode = 27.75
648

 

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