RE digest X - Ligation - RE Digest Y (purif. needed?)

Javier Alzerreca Javier at
Fri Jul 12 17:01:10 EST 1996


We have a fragment cut with EcoRI - both ends. This fragment is pure.
If we proceed to a ligation reaction, (for circularization) do we need 
to purify the DNA from this reaction before a new RE digest with a 
different enzyme? "Will T4 DNA ligase affect the second RE digest?"

Purification is best, but we think we have little DNA and we don't want 
to loose it in a phenol-chloroform extraction. The same may apply for 
other purification techniques.

Thanks for any help.

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