Alice:LabNotes/2007-10-17

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Repeat Exp1: PCR on PGP1 and Hues6 gDNA/cDNA for Sanger sequencing[edit]

Summary of reactions:

Reaction SNP PCR primer Primer location Sample
1(1A) rs3753565 rs3753565_F/rs3753565_R SnpVer2 A1/A12 Hues6 gDNA
2(1B) rs3753565 rs3753565_F/rs3753565_R SnpVer2 A1/A12 Hues6 cDNA
3(1C) rs2295680 rs2295680_F/rs2295680_R SnpVer2 A2/B1 PGP1 gDNA
4(1D) rs2295680 rs2295680_F/rs2295680_R SnpVer2 A2/B1 GM20431 cDNA
5(1E) rs2295680 rs2295680_F/rs2295680_R SnpVer2 A2/B1 GC1F cDNA
6(1F) rs2295680 rs2295680_F/rs2295680_R SnpVer2 A2/B1 GC1EP cDNA
7(1G) rs6103 rs6103_F/rs6103_R SnpVer2 A9/B8 PGP1 gDNA
8(1H) rs6103 rs6103_F/rs6103_R SnpVer2 A9/B8 GM20431 cDNA
9(2A) rs6103 rs6103_F/rs6103_R SnpVer2 A9/B8 GC1F cDNA
10(2B) rs6103 rs6103_F/rs6103_R SnpVer2 A9/B8 GC1EP cDNA
11(2C) rs6103 rs6103_F/rs6103_R SnpVer2 A9/B8 Hues6 gDNA
12(2D) rs6103 rs6103_F/rs6103_R SnpVer2 A9/B8 Hues6 cDNA
13(2E) rs1264899 rs1264899_cF/rs1264899_cR Hues6Ver2c F1/F4 PGP1 gDNA
14(2F) rs1264899 rs1264899_cF/rs1264899_cR Hues6Ver2c F1/F4 GM20431 cDNA
15(2G) rs1264899 rs1264899_cF/rs1264899_cR Hues6Ver2c F1/F4 GC1F cDNA
16(2H) rs1264899 rs1264899_cF/rs1264899_cR Hues6Ver2c F1/F4 GC1EP cDNA

Template: 10ng genomic DNA or 2ng cDNA, use new aliquots of templates/primers/SYBG I diluted in nuclease free H2O.

' one reaction total
Template 1 17
10x Buffer 5 85
10mM dNTP 1 17
10uM forward primer + reverse primer 1 17
50x SYBG I 0.4 6.8
Jump Start Tag DNA Polymerase 1 17
H2O 40.6 690.2

95C 3min -> 35 cycles of (95C 15S -> 58C 30S -> 72C 1min) -> 72C 3min
Result:
The reaction went very well as all the reactions shot up. I will run a gel on these reactions to see if all of them worked.