Haynes Lab:Notebook/CRISPR Editing/2014/07/08: Difference between revisions
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Rene M Davis (talk | contribs) (Autocreate 2014/07/08 Entry for Haynes_Lab:Notebook/CRISPR_Editing) |
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==07//2014== | ==07/08/2014== | ||
Retrying the ligation, won't leave overnight again. | |||
<br> | |||
Annealing gRNA oligos<br> | |||
3ul 100uM top oligo<br> | |||
3ul 100uM bottom oligo<br> | |||
2ul 10x annealing buffer<br> | |||
12ul H2O<br> | |||
20 ul reactions <br><br> | |||
Assemblies<br> | |||
boil and cool method<br> | |||
Aluminum foil lid on the beaker to prevent condensation<br> | |||
Bring ~900 mL water to a boil in a large beaker (on a hot plate). Use a thermometer to check the temperature (>100°C).<br> | |||
Float the oligo mixtures in the boiling water for 10 min. Cover the beaker with aluminum foil to keep the air above the tube warm.<br> | |||
Turn off the heat source and allow the aluminum foil-covered water bath to slowly cool to room temperature (25°C) for several hours.<br><br> | |||
Revision as of 14:57, 8 July 2014
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07/08/2014Retrying the ligation, won't leave overnight again.
3ul 100uM top oligo 20 ul reactions
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