Alice:LabNotes/2008-5-26: Difference between revisions
(New page: Experiment 1: Run a agarose gel on cosmic 3760 probe Image:ZhangLab_2 2008-05-26 14hr 45min.jpeg Result: the gel had consistent result with the PCR curves, which no product was produc...) |
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Experiment 1: Run a agarose gel on cosmic 3760 probe | Experiment 1: Run a agarose gel on cosmic 3760 probe<br> | ||
[[Image:ZhangLab_2 2008-05-26 14hr 45min.jpeg]] | [[Image:ZhangLab_2 2008-05-26 14hr 45min.jpeg]] | ||
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Experiment 2: prepare a cosmic probe with Taq master mix | Experiment 2: prepare a cosmic probe with Taq master mix | ||
Step 1: Real time PCR | Step 1: Real time PCR | ||
reaction system | reaction system x2 | ||
H2O 48.9ul | H2O 48.9ul 97.8 | ||
Taq master mix(2x) 50ul | Taq master mix(2x) 50ul 100 | ||
forward + reverse primer(100uM) 0.5ul | forward + reverse primer(100uM) 0.5ul 1 | ||
SYBR Green (50x) 0.5ul | SYBR Green (50x) 0.5ul 1 | ||
probe template (cosmic 3760) 0.1ul | probe template (cosmic 3760) 0.1ul 0.2 | ||
Total 100ul | Total 100ul 200 | ||
Latest revision as of 16:11, 20 June 2008
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.