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- DNA extraction from Mutation Detection Enhancement (MDE) Gel Stained with Silver Nitrate (Contributed by Mohammad Reza Abbaszadegan et al.)
Single strand conformational polymorphism (SSCP)is the most widely used PCR-based methods for point mutation detection. The abnormal band found by SSCP analysis is normally verified using sequencing. There are many different procedures to recover DNA fragments from the silver stained polyacrylamide gels. However, most of those techniques are time consuming and their recoveries can be low for the Mutation Detection Enhancement (MDE) gel. This is a rapid and efficient method for recovering DNA fragments from MDE gel. In this procedure, the authors have suggested boiling in phenol/ chloroform/isoamylalcohol for elution of DNA from MDE gel that gives high yield of DNA without swelling of the gel. After precipitation of DNA by ethanol and DNA carrier such as glycogenat -70°C, the DNA pellet is rinsed with 75% ethanol to remove contaminations. The yield and quality of resulting DNA from the SSCP variant by this method is sufficient for PCR re-amplification, which can be followed by DNA sequencing to identify mutation. This method can be similarly used for DNA elution from polyacrylamide gels.
Added: Wed Feb 19 2003, Reviews: 0 Write review -
DNA Fragment Purification from Agarose or Acrylamide
(Mike A. Dyer)
For fragments from 200 bp to 10 kb the agarose purification is ideal. For smaller fragments (20 bp to 400 bp) the acrylamide purification is preferred.
http://axon.med.harvard.edu/~cepko/protocol/mike/D...
Added: Tue May 14 2002, Hits: 5754, Reviews: 0 Write review Cached -
Electro-elution of Nucleic Acids from Agarose and Polyacrylamide Gel
(John Roth, Division of Biological Sciences,University of California at Davis)
The nucleic acids contained within the gel slice are electrophoresed out of the gel, funneled down into the v-shaped channel and trapped within a high salt cushion resting at the bottom of the channel.
http://rothlab.ucdavis.edu/protocols/dna-electroel...
Added: Tue May 25 2004, Hits: 5314, Reviews: 0 Write review Cached -
Isolation of DNA from Acrylamide Gels
(Schimenti Lab)
http://lena.jax.org/~jcs/techniques/protocols/acry...
Added: Tue May 14 2002, Hits: 4043, Reviews: 0 Write review Cached -
Oligo Purification on Acrylamide Gels
(Bowtell Lab)
Purification of labeled oligos
http://grimwade.biochem.unimelb.edu.au/bowtell/mol...
Added: Tue May 14 2002, Hits: 1661, Reviews: 0 Write review Cached -
Oligonucleotide purification by PAGE and DEAE
(The Hahn Lab, The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute, Seattle)
http://www.fhcrc.org/labs/hahn/methods/mol_bio_met...
Added: Tue May 14 2002, Hits: 1804, Reviews: 0 Write review Cached