Alice:LabNotes/2008-1-11

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Experiment 1:Preparation of padlock library[edit]

  • Real time PCR setup:
' X16
2X Taq master mix 50 uL 800uL
100 uM forward primer 0.5uL 8uL
100 uM reverse primer 0.5uL 8uL
SYBR Green (50x) 0.5uL 8uL
dH2O 48.2 768
Template (10uM) 0.2 uL 3.2uL
Total 100uL 1600uL
94C 2min
20x (94C 30 sec, 58C 2 min, 72C 1 min)
72C 5 min incubate at 15C forever Note: the PCR is usually stopped 3 cycles after the curve goes to plateau.
Agrose gel result (1-12-08): 
File:ZhangLab 2 2008-01-12 15hr 00min.jpeg
The bright bands on the bottom are the ladders for both columns (3uL ladder, 1 uL TBE buffer, and 1 uL of loading dye),
and the rest of the wells contains sample 1-16 (4uL sample and 1uL of loading dye).
The bands on the left column (first 8 samples) don't good intensity, which is possibly caused by not well-mixed PCR reagents in the tube.