Help:Protocols/Restriction Digest
Revision as of 22:20, 27 September 2010 by Vinoo (Talk | contribs) (New page: 1. Add 500ng of DNA to be digested, and adjust with dH20 for a total volume of 42.5ul.<br> 2. Add 5ul of NEBuffer 2 to the tube.<br> 3. Add 0.5ul of BSA to the tube.<br> 4. Add 1ul of you...)
1. Add 500ng of DNA to be digested, and adjust with dH20 for a total volume of 42.5ul.
2. Add 5ul of NEBuffer 2 to the tube.
3. Add 0.5ul of BSA to the tube.
4. Add 1ul of your first enzyme.
5. Add 1ul of your second enzyme.
6. There should be a total volume of 50ul. Mix well and spin down.
7. Incubate the restriction digest at 37C for 30min, and then 80C for 20min to heat kill the enzymes.
8. Run a portion of the digest on a gel, to check that both plasmid and part length are accurate.