RE digest X - Ligation - RE Digest Y (purif. needed?)
Javier at mendel.usu.edu
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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