Colony-PCR: Difference between revisions
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*0,2 µl Primer rev | *0,2 µl Primer rev | ||
*0,04 µl Taq Polymerase | *0,04 µl Taq Polymerase | ||
* | *8,36 µl Water | ||
==PCR Program== | ==PCR Program== | ||
Latest revision as of 23:12, 29 August 2013
Instead of plasmid or genomic DNA, you can use a colony from the agar plate as template DNA. You should use a sterile toothpick, pick into the colony and spread it in the PCR tube.
For a 10 µl reaction (Taq Polymerase Peqlab):
- 1 µl Buffer S
- 0,2 µl dNTP's (10 mM)
- 0,2 µl Primer fw
- 0,2 µl Primer rev
- 0,04 µl Taq Polymerase
- 8,36 µl Water
PCR Program
- 95°C 5 min
35 times the following cycle
- 95°C 1 min
- 55°C 30 sek. (°C depends on your primers)
- 72°C 1 min/kb
cycle end
- 72°C 10 min
- 10°C hold
