Kun:LabNotes/ASE/2007-11-29

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Exp. 1 Perform PCR/sequencing on GM20431/GC1F/GC1EP/Hues6 cDNAs[edit]

I've done PCR/sequencing on the PGP1VER2 primer set, and some of the reactions failed in the first round of experiment. I'm repeating the PCR on the failed reactions. I will use primers from the master plate.

PCR tube Primer SNP Template
1 A1 rs1045216 GC1F
2 B1 rs11264743 GM20431
3 B1 rs11264743 GC1F
4 B1 rs11264743 GC1EP
5 B1 rs11264743 Hues6 c
6 F1 rs1869214 GM20431
7 F1 rs1869214 GC1F
8 F1 rs1869214 GC1EP
9 G1 rs2271504 GC1F
10 G1 rs2271504 GC1EP
11 A2 rs2295230 GM20431
12 A2 rs2295230 GC1F
13 A2 rs2295230 GC1EP
14 C2 rs2861158 GM20431
15 C2 rs2861158 GC1F
16 C2 rs2861158 GC1EP
17 E2 rs3829655 GC1F
18 E2 rs3829655 GC1EP
19 E2 rs3829655 Hues6 c
20 F2 rs6664 Hues6 c
21 G2 rs7198 Hues6 c
22 H3 rs12436 GM20431
23 H3 rs12436 GC1F
24 H3 rs12436 GC1EP
                      One       25x       
  Template            1ul       each     
  10X buffer          5ul      125ul    
  25mM MgCl2          3ul       75ul
  10mM dNTP           1ul       25ul
  50X SYBG I        0.4ul       10ul
  10uM Primers        1ul       each
  iTaq             0.25ul     6.25ul
  H2O             38.35ul      960ul

94C 3min -> 40 cycles of (94C 30S -> 58C 30S -> 72C 1min) -> 72C 3min on Chromo4.