Cloning PCR fragment into the expression vector

Michael L. Sullivan mlsulliv at facstaff.wisc.edu
Thu Mar 7 14:42:26 EST 2002


>What about purity of agarose?  What brands and types work well w/ gel
>purification and downstream ligation of DNA?
>
>These issues have impacted my work in the past.
>

For the past 6 or 7 years, I've mostly been using Gibco (no affiliation)
agarose.  Most often, for purifying fragments, I use "band capturing" on
S&S NA45 paper and subsequent elution and precipitation prior to inclusion
in the ligation.  Sometimes, I will also carry out a ligation with fragment
cut out of low melting agarose, melted in water, and used directly in the
ligation reaction, also Gibco agarose.

Hope this helps.  Let me know if you need more information.

Mike

Michael L. Sullivan, Ph.D

U.S. Dairy Forage Research Center
1925 Linden Drive West
Madison WI, 53706

(608) 264-5144 Phone
(608) 264-5147 Fax

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