Alice:LabNotes/2008-5-26

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Experiment 1: Run a agarose gel on cosmic 3760 probe

File:ZhangLab 2 2008-05-26 14hr 45min.jpeg

Result: the gel had consistent result with the PCR curves, which no product was produced from the amplification. However, the gel should have some primer dimers, but none is shown. This is possibly caused by not loading enough samples. The first lane is the ladder, which had 0.5 ul of 100 bp ladder, diluted with 3.5 ul H2O, and 1 ul of 5% TBE dye. The rest of the lanes had 4 ul of PCR product, and 1 ul of 5% TBE dye.

Experiment 2: prepare a cosmic probe with Taq master mix

Step 1: Real time PCR

 reaction system                                 x2   
 H2O                              48.9ul        97.8         
 Taq master mix(2x)                 50ul        100   
 forward + reverse primer(100uM)   0.5ul        1              
 SYBR Green (50x)                  0.5ul        1 
 probe template (cosmic 3760)      0.1ul        0.2
 Total                             100ul        200


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.