Double Digest using enzymes from diff companies - (May/04/2009 )

I need to do a double digest with XhoI from Takara and EcoRI from NEB. What do I do about buffer compatibility? Will the EcoRI from NEB work with EcoRI buffer from Takara (will also work with XhoI)?

I'm crossing my fingers.... haha

Thanks!

Look at the buffer composition and compatibility. You should find it easily on the NEB website.

ChrissyFL on May 4 2009, 08:05 PM said:

Takara recommend their buffer H for both enzymes. NEB suggests their unique "EcoR1 buffer"

Thus, do your double digest in either Buffer H or NEB "EcoR1 buffer".

This will work fine as the buffers have very similar salt concentrations.

Hope this helps

Hey

If you are not satisfied with double digestions, just digest with the first enzyme, precipitate and digest with the other sequentially. Thats what I do for enzymes from different companies. Better delay at this stage rather than repeating the whole cycle. However, if buffers are compatible there is no reason why double digestion should not work.

Best,
TC

Thanks for the suggestions!