PrbbBB:colony pcr v1

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Revision as of 07:55, 26 February 2009 by Raik (Talk | contribs)
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Contents

Overview

Materials

Procedure

I PCR reaction

11µl single reaction 60xMaster 110xMaster
H2O 7.5µl
10x Amplitaq buffer 1.1µl
10mM dNTP 0.22µl
VF2 primer 10 µM 0.55µl
VR primer 10 µM 0.55µl
AmpliTaq 5U/µl 0.1µl
template DNA 1µl -- --
PCR Program
30"@98°C
5x (10"@98°C; 15"@Ta; text@72°C);
25x (10"@98°C; text@72°C);
10'@72°C
∞ 4°C
  • extension time text = (kb insert length) × 25"
  • annealing temperature Ta = (primer annealing) + 3°C

II Agarose Gel

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

raik: no comment


References

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