Alice:LabNotes/2007-11-7: Difference between revisions

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Template: 10ng genomic DNA or 2ng cDNA, use new aliquots of templates/primers/SYBG I diluted in nuclease free H2O.
Template: 10ng genomic DNA or 2ng cDNA, use new aliquots of templates/primers/SYBG I diluted in nuclease free H2O.
' one reaction total
' one reaction total
Template 1 17
Template 1 17

Revision as of 22:29, 7 November 2007

The sequencing result showed that Reaction #1, 2, 3, 12, 14, 16 didn't work as expected, as they had too much overlap, so the experiment on these reactions will be repeated.

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

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

12(2D) rs6103 rs6103_F/rs6103_R SnpVer2 A9/B8 Hues6 cDNA

14(2F) rs1264899 rs1264899_cF/rs1264899_cR Hues6Ver2c F1/F4 GM20431 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