Extracting DNA from low-melt agarose gels
sdorazio at miasun.med.miami.edu
Thu Jul 8 10:51:07 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
We only phenol extract once, and have had no problems with
this relatively simple method:
1. Cut out your band, and melt at 70 C.
2. Add an equal volume of prewarmed (to 37 C) phenol;
3. Vortex; quick freeze in dry ice/EtOH bath
4. While still frozen, spin in microfuge at 4 C for 5
5. EtOH ppt aqueous layer. The agarose should be at
the bottom of the tube.
Sarah D'Orazio sdorazio at miasun.med.miami.edu
Univ. of Miami Sch. of Med.
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