Cloning Protocol: Difference between revisions
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Step 1: Phusion PCR | |||
=== Step 1: Phusion PCR === | |||
Do a 200 uL reaction for the insert (2 PCR tubes with 100 uL) | Do a 200 uL reaction for the insert (2 PCR tubes with 100 uL) | ||
<!-- <mediaplayer width='500' height='300'>http://www.bradleymonk.com/w/images/c/cc/Cloning1.mp4</mediaplayer> --> | |||
{{#widget:Html5media|url=http://www.bradleymonk.com/w/images/c/cc/Cloning1.mp4|width=400}} | |||
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Revision as of 10:31, 11 January 2016
Step 1: Phusion PCR
Do a 200 uL reaction for the insert (2 PCR tubes with 100 uL)
{{#widget:Html5media|url=http://www.bradleymonk.com/w/images/c/cc/Cloning1.mp4%7Cwidth=400}}
| 10 uM forward primer | 10 uL | |
| 10 uM reverse primer | 10 uL | |
| 2x phusion master mix (MM) | 100 uL | |
| 10 ng total template | 1 uL total | |
| H2O up to 200 uL | variable - top to 200 uL |