Haynes Lab:Notebook/CRISPR Editing/2016/03/12: Difference between revisions

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==03/11/2016==
==03/12/2016==
Colony PCR of colonies on ligation plates from Saturday. Primers P220/221 will give a 1240bp band if GFP is inserted. will give a 439bp for backbone only. Will be slightly longer if the backbone was only cut once. Used 2x taq MM. NEB suggests an annealing T of 51°C with Taq, will do 55°C to try to avoid lots of off target binding to gDNA. <br>Ran on gel
Colony PCR of colonies on ligation plates from Saturday. Primers P220/221 will give a 1240bp band if GFP is inserted. will give a 439bp for backbone only. Will be slightly longer if the backbone was only cut once. Used 2x taq MM. NEB suggests an annealing T of 51°C with Taq, will do 55°C to try to avoid lots of off target binding to gDNA. <br>Ran on gel
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Revision as of 17:51, 12 March 2016

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03/12/2016

Colony PCR of colonies on ligation plates from Saturday. Primers P220/221 will give a 1240bp band if GFP is inserted. will give a 439bp for backbone only. Will be slightly longer if the backbone was only cut once. Used 2x taq MM. NEB suggests an annealing T of 51°C with Taq, will do 55°C to try to avoid lots of off target binding to gDNA.
Ran on gel

Miniprepped g031 in pX330g

Ran gDNA extractions on gel since I left them lysing at 56deg for 18 hours

Moved cells to T25 and T75 flask