Kun Zhang LabNote Oct week1: Difference between revisions

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(New page: ==Thursday 2007-10-04== ===Thu.Exp1: PCR on PGP1 and Hues6 gDNA/cDNA for Sanger sequencing=== Summary of reactions: {| {{table}} | align="center" style="background:#f0f0f0;"|'''Reaction'''...)
 
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Template: 10ng genomic DNA or 5ng cDNA
Template: 10ng genomic DNA or 5ng cDNA
{| {{table}}
{| {{table}}
| align="center" style="background:#f0f0f0;"|''' '''
| align="center" style="background:#f0f0f0;"|''''''
| align="center" style="background:#f0f0f0;"|'''one reaction'''
| align="center" style="background:#f0f0f0;"|'''one reaction'''
| align="center" style="background:#f0f0f0;"|'''total'''
| align="center" style="background:#f0f0f0;"|'''total'''
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| 10x Buffer||5||85
| 10x Buffer||5||85
|-
|-
| 25mM dNTP||0.4||6.8
| 50mM MgCl2||1.5||25.5
|-
| 10mM dNTP||1||17
|-
|-
| 10uM forward primer||1||17
| 10uM forward primer||1||17
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| 50x SYBG I||0.4||6.8
| 50x SYBG I||0.4||6.8
|-
|-
| iTaq||0.5||8.5
| iTaq||0.25||4.25
|-
| H2O||38.85||660.45
|-
|-
|  
|  

Revision as of 19:02, 4 October 2007

Thursday 2007-10-04

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

Summary of reactions:

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

Template: 10ng genomic DNA or 5ng cDNA

' one reaction total
Template 1 17
10x Buffer 5 85
50mM MgCl2 1.5 25.5
10mM dNTP 1 17
10uM forward primer 1 17
10uM reverse primer 1 17
50x SYBG I 0.4 6.8
iTaq 0.25 4.25
H2O 38.85 660.45

95C 3min -> 35 cycles of (95C 15S -> 58C 30S -> 72C 1min) -> 72C 3min