extracting dna from acrylamide gels

[Alexander Dobrovic]

We use the bandstab technique of Wilton (1997). Bands are stabbed with a
yellow tip which is then placed in 10ul of PCR grade water. 1ul is used for
PCR.

This saves a lot of time and can be applied to any bands in a polycrylamide
gel.

Bianco T, Hussey D, Dobrovic A.  Methylation-sensitive single-strand
conformation analysis (MS-SSCA): A rapid method to screen for and analyze
methylation.  Hum Mutat. 1999;14(4):289-93.

Wilton SD, Lim L, Dye D, Laing N.   Bandstab: a PCR-based alternative to
cloning PCR products. Biotechniques. 1997 Apr;22(4):642-5.


>I'm looking for advice on extracting DNA bands from acrylamide gels.  My
>goal is to recover enough DNA for a few PCR reactions. The acrylamide gels
>are stained in EtBr and were made with FMC's Long Ranger mix.
>
>I've been using the ammonium acetate elution protocol from Maniatis, but
>I'm not finding it to be very reliable in my hands and it would be nice to
>find a procedure that doesn't require an o/n incubation.
>
>I'm wondering whether there's a simpler and quicker way that will work for
>PCR.  Do acrylamide or EtBr inhibit PCR?  Could I just elute the gel slice
>in water to avoid having to get rid of the ammonium acetate?  Could I just
>freeze (or boil) the gel slice?  Any thoughts or anecdotes are welcome.
>
>
>
>---


Alexander Dobrovic, Ph.D.
Chief Medical Scientist/Dept of Haematology-Oncology
Affiliate Senior Lecturer/University of Adelaide Dept of Medicine
The Queen Elizabeth Hospital
Woodville, SA 5011, Australia

Tel  61-8-8222 7676
Fax  61-8-8222 6046
alexander.dobrovic at adelaide.edu.au



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