Alice:LabNotes/2008-1-11
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Experiment 1:Preparation of padlock library
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.