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.