User:Stuart McKellar/Notebook/Bird Sex Testing/2013/01/19: Difference between revisions
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== | ==Preliminary results generation== | ||
* | * I am getting results that Iam confident are usable. This is by comparing them to the positive control samples and getting ratios that are correct. | ||
I will be doing a PCR with P2/P8 primers and then comparing against positive controls with ALL SAMPLES (except loris). Then I will repeat this with the 2550F primer pair at 55°C and comparing all these results. If I can't get consistency, I will order the NP MP primer set. | |||
Method: | |||
black pen - P2 - 60C | |||
blue - 2550F - 55C | |||
29 samples | |||
30 samples master mix | |||
Tube | |||
*1ul F primer | |||
*1ul R primer | |||
*2.0ul dNTPs | |||
*0.5u Taq | |||
*1ul DNA | |||
*2.5ul 10x buffer | |||
*1.5ul MgCl<sub>2</sub> | |||
*16ul H<sub>2</sub>O | |||
Master mix | |||
*30ul F primer yes | |||
*30ul R primer yes | |||
*60ul dNTPs yes | |||
*3ul Taq (0.5u per tube) | |||
*75ul 10x buffer | |||
*45ul MgCl<sub>2</sub> | |||
*480ul H<sub>2</sub>O | |||
added 35ul oil | |||
3min initial melt | |||
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Revision as of 23:26, 18 January 2013
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Preliminary results generation
I will be doing a PCR with P2/P8 primers and then comparing against positive controls with ALL SAMPLES (except loris). Then I will repeat this with the 2550F primer pair at 55°C and comparing all these results. If I can't get consistency, I will order the NP MP primer set. Method: black pen - P2 - 60C blue - 2550F - 55C 29 samples
Tube
added 35ul oil 3min initial melt | |
