Kun:LabNotes/ASE/2008-6-27

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Sanger confirmation of ASE in PGP1[edit]

PCR[edit]

  • Template:
    • PGP1L: 137ng/ul, 06/03/08
    • PGP1K: 80ng/ul, 06/03/08
    • PGP1F: 101ng/ul, 06/24/08
  • Primers: PGP1L-ASE-conf (06/25/08), 48 pairs.
                                x 50 x 3
   Template:           4ng   
   2X TaqMaster mix:  15ul      750
   5uM primer mix:   1.5ul      each
   50X SYBGI:        0.1ul        5
   H2O                13ul      650
   94C 2min -> 35x (94C 30sec -> 57C 30sec -> 72C 30sec) -> 72C hold.
  PGP1L/PGP1K
  File:2008-06-27-cDNA-PCR.png
1 2 3 4 5 6 7 8 9 10 11 12
A X X X X X X X X X X X O
B O X X X X X O X X X X X
C X X X X X X X X X X X X
D X X O O O X O O X X X X
E X O O X X X O O X X X X
F X X X X X X X O X X X X
G O X O X O X X X X X O X
H O X O X X O X X X X X X
  PGP1F  (A=====>H)
  File:2008-06-30-PGP1F-cDNA-PCR.png
  • Also perform PCR on cDNA from PGP1L made on 12/31/07.
  Layout of the plate shipped to Agencourt on 06/30/08
  File:PGP1L-conf-submitted-sample-layout-06302008.png