Team:EPF-Lausanne/Protocols/PCR

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Primers
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==Primers==
Make primers pair mix with 3µM concentration (total).
Make primers pair mix with 3µM concentration (total).
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'''<big>Cycles</big>'''
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==Cycles==
''Program PCR: count 1min/kb''
''Program PCR: count 1min/kb''
:94°C – 120’’
:94°C – 120’’
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:94°C – 45’’ \{
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:\{94°C – 45’’  
:56°C – 45’’          → 30x
:56°C – 45’’          → 30x
:68°C – 120’’
:68°C – 120’’
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:72°C – 420’' \}
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:\}72°C – 420’'  
:10°C – ∞
:10°C – ∞

Revision as of 07:19, 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




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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