Alice:LabNotes/2007-11-14: Difference between revisions

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[[Image:ZhangLab_2 2007-11-14 17hr 10min.jpg]]
[[Image:ZhangLab_2 2007-11-14 17hr 10min.jpg]]


Also, I did PCR on sample #14 with newly designed primers rs1264899 along with template GC1EP, GC1F, GM20431, PGP1. <br>
<br>
<b>Experiment 2: PCR on sample #14 with newly designed primers rs1264899 </b> <br>
Summary of reactions:
Summary of reactions:
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Revision as of 00:13, 1 December 2007

I ran a gel of the PCR product from repeated experiment on sample #1,2,3,12,14,16. The gel image shows that all the reactions worked fine, so they can be prepared for sequencing. Eventually, the post PCR products are prepared with EXOSap.
File:ZhangLab 2 2007-11-14 17hr 10min.jpg


Experiment 2: PCR on sample #14 with newly designed primers rs1264899
Summary of reactions:

Reaction SNP PCR primer Sample
1 rs1264899 rs1264899_F/rs1264899_R GC1EP
2 rs1264899 rs1264899_F/rs1264899_R GC1F
3 rs1264899 rs1264899_F/rs1264899_R GM20432 cDNA
4 rs1264899 rs1264899_F/rs1264899_R PGP1 gDNA
' 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

I forgot the reaction was only for 4 samples, therefore the left over master mix is stored in -20 fridge and saved for later use.