Brandon:Protocols/taq amplification2: Difference between revisions
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==taq amplification | ==Standard PCR cycling methods for different enzymes/procedures== | ||
*KAPA SYBR | |||
12.5 uL KAPA SYBR | |||
2 uL primers, 1 uL F, 1 uL R | |||
5.5 uL H2O | |||
5 uL DNA template | |||
KAPA SYBR cycles: | |||
98C 3min, (98C for 30s, 60C for 30s, 72C for 1 min) X35, 72C for 5 min, 4C forever | |||
*taq amplification | |||
**also used for colony PCR, pick colony and put directly into the reaction | |||
used taq2x for amplification, X35 cycles | used taq2x for amplification, X35 cycles | ||
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taq2x cycles: | taq2x cycles: | ||
95C 5min, (95C for 30s, 60C for 30s, 72C for 1 min) X35, 72C for 5 min, 4C forever | 95C 5min, (95C for 30s, 60C for 30s, 72C for 1 min) X35, 72C for 5 min, 4C forever | ||
*KAPA2G Robust |
Latest revision as of 19:13, 20 May 2012
Standard PCR cycling methods for different enzymes/procedures[edit]
- KAPA SYBR
12.5 uL KAPA SYBR 2 uL primers, 1 uL F, 1 uL R 5.5 uL H2O 5 uL DNA template KAPA SYBR cycles: 98C 3min, (98C for 30s, 60C for 30s, 72C for 1 min) X35, 72C for 5 min, 4C forever
- taq amplification
- also used for colony PCR, pick colony and put directly into the reaction
used taq2x for amplification, X35 cycles
12.5 uL taq2X 2 uL primers, 1 uL F, 1 uL R 5.5 uL H2O 5 uL DNA template taq2x cycles: 95C 5min, (95C for 30s, 60C for 30s, 72C for 1 min) X35, 72C for 5 min, 4C forever
- KAPA2G Robust