Alice:LabNotes/2008-5-12

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Revision as of 21:19, 12 May 2008 by >Zsakura2 (New page: Experiment 1: Padlock Library preparation for probe Cosmic Step 1: Real time PCR reaction system x 16 H2O 48 ul ...)
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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.