User:Stuart McKellar/Notebook/Bird Sex Testing/2012/12/01
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Current revision (01:31, 1 December 2012) (view source) (→higher annealing temp) |
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total volume in each is 25μL. | total volume in each is 25μL. | ||
| - | PCR: 94°C for 2m30s, 35 cycles of: 95°C 30s, 65°C for 30s, 72°C for 30s; final elongation 72°C for 5 mins. | + | PCR: 94°C for 2m30s, 35 cycles of: 95°C 30s, 65°C for 30s, 72°C for 30s; final elongation 72°C for 5 mins( actually went 10 mins by accident). |
results to be determined via agar gel electrophoresis on 1% gel stained using gelgreen. | results to be determined via agar gel electrophoresis on 1% gel stained using gelgreen. | ||
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higher annealing tempAim: test the p2/p8 and 2550/2178 primers at 65°C. Method: Sample 1:
Sample 2:
Negative control - no DNA (1ul h2O, 2550/2178 primers.) Master mix (5x):
total volume in each is 25μL. PCR: 94°C for 2m30s, 35 cycles of: 95°C 30s, 65°C for 30s, 72°C for 30s; final elongation 72°C for 5 mins( actually went 10 mins by accident). results to be determined via agar gel electrophoresis on 1% gel stained using gelgreen.
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