Extracting DNA from low-melt agarose gels
drmax at casbah.acns.nwu.edu
Fri Jul 9 06:54:09 EST 1993
In article <01H0AHG3K9OY8WW9E0 at Butler.EDU> KARN at Butler.EDU writes:
>I wish to use low-melt agarose gels to clean up a PCR product of about 1 kb
>(+/- 0.2 kb) but would like to avoid the multiple extraction procedure
>described in Maniatis. My goal is to sequence the product so it must be
>relatively clean. Anybody got an improvement on the Maniatis technique
>or must I haul out the phenol?
>KARN at BUTLERU.edu
I use a product called Qiaex (sp?) from Qiagen. You run the DNA in regular
agarose, cut out the band and combine it with a DNA binding matrix and a
buffer that "melts" the agarose at 37 - 50oC in about 5m. Spin this down
and wash 2x with the next two buffers. Resuspend in 20ul of water and use.
I've had no problems using this directly for ligations and the yeild is at
least as good if not better than any other method that I've used.
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