Haynes Lab:Notebook/CRISPR Editing/2014/07/08

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07/08/2014

Retrying the ligation, won't leave overnight again.
Annealing gRNA oligos

3ul 100uM top oligo
3ul 100uM bottom oligo
2ul 10x annealing buffer
12ul H2O

20 ul reactions

Assemblies
boil and cool method
Aluminum foil lid on the beaker to prevent condensation
Bring ~900 mL water to a boil in a large beaker (on a hot plate). Use a thermometer to check the temperature (>100°C).
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.
Turn off the heat source and allow the aluminum foil-covered water bath to slowly cool to room temperature (25°C) for several hours.



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