Team:EPF-Lausanne/Protocols/PCR

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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

Cycles

Program PCR: count 1min/kb

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