Alice:LabNotes/2008-5-12
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Experiment 1: Padlock Library preparation for probe Cosmic
Step 1: Real time PCR
reaction system x 16 H2O 48 ul ul JumpStart Taq DNA polymerase 0.5 ul 8 ul 10X jumpstart Taq buffer 10 ul 160 ul dNTP primer mix: Ap2V4 + Ap1V4IU 1.0 ul 16 ul SYBR reen (50x) 0.5 ul 8 ul probe template (CPGCES 20uM) 0.1 ul 1.6 ul Total 100 ul 1600 ul
Reaction program:
94c 2min -> 94c 30sec -> 60C 2min -> 72C 1min-> Plate read -> (94c 30sec -> 60C 2min -> 72C 1min)x 20 cycles -> 72C 5min -> 15C hold.
Primer sequence: forward primer Ap1V4IU and reverse primer Ap2V4
Note: 32 tubes were prepared instead of 16.