PrbbBB:colony pcr v1

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(New page: Back to all protocols ==Overview== ==Materials== *96-well PCR plate *AmpliTaq DNA Polymerase 5 U/µl (Roche) *AmpliTaq Buffer 10 x *dNTP mix 10mM each nucleotide ...)
(Materials)
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*ddH2O
*ddH2O
*primers:
*primers:
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**BBa_G00100 (VF2) [http://partsregistry.org/wiki/index.php/Part:BBa_G00100 MIT Registry] [http://brickit.crg.es/registry/biobrick/5/ CRG-internal registry]
+
**[http://partsregistry.org/wiki/index.php/Part:BBa_G00100 BBa_G00100 (VF2)] [http://brickit.crg.es/registry/biobrick/5/ CRG-internal registry]
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**BBa_G00101 (VR)  [http://partsregistry.org/wiki/index.php/Part:BBa_G00101 MIT Registry] [http://brickit.crg.es/registry/biobrick/6/ CRG-internal registry]
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**[http://partsregistry.org/wiki/index.php/Part:BBa_G00101 BBa_G00101 (VR)] [http://brickit.crg.es/registry/biobrick/6/ CRG-internal registry]
==Procedure==
==Procedure==

Revision as of 07:50, 26 February 2009

Back to all protocols

Contents

Overview

Materials

Procedure

II PCR reaction

100µl single reaction 3.5xMaster 10xMaster
H2O 76µl 266 760
5x HF Buffer 20µl 70 200
10mM dNTP 2µl 7 20
FW primer 100 µM 0.5µl 1.75 5
RV primer 100 µM 0.5µl 1.75 5
Phusion 1µl 3.5 10
template DNA 0.1µl -- --
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

Contact

or instead, discuss this protocol.

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