Extracting DNA from low-melt agarose gels

Sarah Dorazio 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?
>Bob Karn
>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
	minutes
	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.
	Miami, Florida
B

	4.	W



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