Team:EPF-Lausanne/Protocols/PCR

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We used '''2''' tubes of PCR of 25µl each.
==Cycles==
==Cycles==

Revision as of 07:25, 4 September 2009



Contents

PCR Protocol



Primers

Make primers pair mix with 3µM concentration (total).

Reaction mix
Plasmid 0.5ul
Primers pair (3µM) 1ul
Thermo POL buffer 2.5µl
dNTP 0.5 µl
Taq EPFL polym 0.25 µl
H20 (MQ) 20.25 µl
Total 24.5 µl


We used 2 tubes of PCR of 25µl each.

Cycles

Program PCR: count 1min/kb

94°C – 120’’
94°C – 45’’        |
56°C – 45’’        |    → 30x
68°C – 120’’      |
72°C – 420’'      |
10°C – ∞



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