best purification of 186bp DNA fragment from agarose

pnh at ncifcrf.gov pnh at ncifcrf.gov
Mon Nov 21 19:50:37 EST 1994


Robin Colgrove (robin at alumni.caltech.edu) wrote:

| I am cloning a small (186bp) PCR product. Usually I clone larger fragments
| and purify the gel fragments with "geneclean." Their package insert
| suggests not using it for less than 200bp. In the old days, I used all
| the standard tricks: Low melt agarose, electroelution, mashing up the 
| gel slice, etc. I was just wondering what people tihink is the best way
| to go about it these days. Thanks.

I say go ahead with GeneClean. I've heard from many people that the recovery
is highly dependent on the specific sequence. I was able to subclone a 35
bp fragment eluted from glassmilk on the first try, but I started with alot
of DNA. For other ideas, you could look at the review article I recently wrote.

@article{Hengen1994Septibs, 
author = "P. N. Hengen",
title = "Methods and Reagents - Recovering {DNA} from agarose gels",
journal = "Trends in Biochemical Sciences",
volume = "19",
number = "9",
pages = "388-389",
month = "September",
year = "1994"}

-Paul.

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