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(New page: ===Exp1: PCR on PGP1 and Hues6 gDNA/cDNA for Sanger sequencing=== Summary of reactions: {| {{table}} | align="center" style="background:#f0f0f0;"|'''Reaction''' | align="center" style="bac...)
 
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===Exp1: PCR on PGP1 and Hues6 gDNA/cDNA for Sanger sequencing===
*[[Alice:Single cell tracing and RNA-seq Lab Notes|Single cell RNA-seq Lab Notes]]
Summary of reactions:
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Reaction'''
| align="center" style="background:#f0f0f0;"|'''SNP'''
| align="center" style="background:#f0f0f0;"|'''PCR primer'''
| align="center" style="background:#f0f0f0;"|'''Primer location'''
| align="center" style="background:#f0f0f0;"|'''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.
{| {{table}}
| align="center" style="background:#f0f0f0;"|''''''
| align="center" style="background:#f0f0f0;"|'''one reaction'''
| align="center" style="background:#f0f0f0;"|'''total'''
|-
| Template ||1||17
|-
| 10x Buffer||5||85
|-
| 25mM MgCl2||3||51
|-
| 10mM dNTP||1||17
|-
| 10uM forward primer||1||17
|-
| 10uM reverse primer||1||17
|-
| 50x SYBG I||0.4||6.8
|-
| thermo start Taq||0.25||4.25
|-
| H2O||37.35||634.95
|-
|
|}


95C 3min -> 35 cycles of (95C 15S -> 58C 30S -> 72C 1min) -> 72C 3min <br>
*[[Alice:Whole Genome Bisulfite Sequencing Lab Notes|Whole Genome Bisulfite Sequencing Lab Notes]]
 
*[[Alice:Exome Sequencing Analysis|Exome sequencing analysis]]
 
*[[Alice:Exome Sequencing Lab Notes-2|Exome sequencing Lab Notes mid-2012 to 2014]]
 
*[[Alice:LabNotes|Exome sequencing Lab Notes 2007 to mid-2012]]
 
*[[Alice:iPSC Reprogramming Lab Notes|hiPSC Reprogramming Lab Notes]]
 
*[[Alice:Lab Meeting Slides|Lab Meeting Slides]]
 
*[[Alice:Sample Record|Sample Tracking]]
 
*[[Alice:Sequencing Sample Record|Sequencing Sample Record]]
 
*[[Alice:Protocols|Protocols]]

Latest revision as of 17:46, 7 March 2016