User:Stuart McKellar/Notebook/Bird Sex Testing/2013/01/19
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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 02:26, 19 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 | |
