Alice:LabNotes/2007-11-14
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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.