Smolke:Protocols/Digests: Difference between revisions
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(New page: {{Smolke_Top]] ==Josh's double digest mix== *1.5ug vector *2uL 10x buffer *0.2uL 100x BSA *0.5uL each enzyme *Water to 20uL Run overnight at 37C. Adjust this mix depending on whether yo...) |
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==Josh's double digest mix== | ==Josh's double digest mix== |
Revision as of 12:09, 15 October 2009
Josh's double digest mix
Run overnight at 37C. Adjust this mix depending on whether you're making a master mix. For one reaction, I up the BSA to 0.5uL (since I don't trust pipetting 0.2uL). For a master mix, you can judiciously reduce the amount of enzyme (<0.5uL/rxn) and adjust the relative enzyme amounts depending on the enzyme concentration (so use more an an enzyme at 5 U/uL and less of one at 20 U/uL). CIP
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