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
|