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Team:Calgary/Notebook/Protocols/KapaPCR
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<li>0.5µL Kapa Hifi DNA polymerase</li> | <li>0.5µL Kapa Hifi DNA polymerase</li> | ||
<li>10 to 50ng of DNA</li> | <li>10 to 50ng of DNA</li> | ||
| - | <li> | + | <li>ddH<sub>2</sub>O (deionized water) up to 25µL</li> |
</ul> | </ul> | ||
<h2>Protocol</h2> | <h2>Protocol</h2> | ||
Latest revision as of 00:19, 28 September 2013
Kapa PCR
Kapa PCR (for DNA constructions)
Mix (per reaction)
- 5.0µL 5x Kapa Hifi Buffer
- 0.75µL 10mM dNTPs
- 0.75µL Primer Forward
- 0.75µL Primer Reverse
- 0.5µL Kapa Hifi DNA polymerase
- 10 to 50ng of DNA
- ddH2O (deionized water) up to 25µL
Protocol
- Add the mix to each tube
- Set the following program in the thermocycler:
- Stage 1 (1 cycle): 95°C for 2 to 5 min
- Stage 2 (30 cycles): 98°C for 20 sec, 55°C for 15 sec, 72°C for 1min/kb
- Stage 3 (1 cycle): 72°C for 1 to 5 min
- Hold at 4°C
- Run the products in a agarose gel
