Difference between revisions of "LongAmp PCR"
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(Created page with "== Preparation == 1. 0.6uL 10mM dNTPs 2. 1uL 1ng template 3. 1uL 10uM forward primer 4. 1uL 10uM reverse primer 5. 1uL LongAmp Taq Polymerase 6. 4uL LongAmp Buffer 7. 1...") |
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| Line 11: | Line 11: | ||
5. 1uL LongAmp Taq Polymerase | 5. 1uL LongAmp Taq Polymerase | ||
| − | 6. 4uL LongAmp Buffer | + | 6. 4uL 5X LongAmp Buffer |
7. 11.4uL dH20 | 7. 11.4uL dH20 | ||
| Line 28: | Line 28: | ||
94ºC 30 seconds | 94ºC 30 seconds | ||
| − | + | 55ºC 30 seconds (Or a different annealing temperature) | |
| − | + | 65ºC 10 minutes (Or 1 minute per 1000bp) | |
'''Stage 3:''' | '''Stage 3:''' | ||
| − | + | 65ºC 10 minutes | |
Latest revision as of 18:55, 10 July 2014
Preparation
1. 0.6uL 10mM dNTPs
2. 1uL 1ng template
3. 1uL 10uM forward primer
4. 1uL 10uM reverse primer
5. 1uL LongAmp Taq Polymerase
6. 4uL 5X LongAmp Buffer
7. 11.4uL dH20
Programed PCR Protocol
Stage 1:
94ºC 5 minutes
Stage 2:
30 cycles
94ºC 30 seconds
55ºC 30 seconds (Or a different annealing temperature)
65ºC 10 minutes (Or 1 minute per 1000bp)
Stage 3:
65ºC 10 minutes
