Protocols
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Denaturing Gradient Gel Electrophoresis (DGGE)
(Donis-Keller lab)
Denaturing gradient gels are used to detect non-RFLP polymorphisms. The small (200-700 bp) genomic restriction fragments are run on a low to high denaturant gradient acrylamide gel; initially the fragments move according to molecular weight, but as they progress into higher denaturing conditions, each (depending on its sequence composition) reaches a point where the DNA begins to melt. The partial melting severely retards the progress of the molecule in the gel, and a mobility shift is observed. It is the mobility shift which can differ for slightly different sequences (depending on the sequence, as little as a single bp change can cause a mobility shift). Alleles are detected by differences in mobility.
http://hg.wustl.edu/hdk_lab_manual/dgge/dgge1.html
Added: Tue May 14 2002, Hits: 1531, Reviews: 0 Write review Cached -
Denaturing Gradient Gel Electrophoresis (DGGE)
(electrophoresis.net)
If you are new to DGGE, this will introduce you to this technique.
http://www.electrophoresis.net/tech_dgge.asp
Added: Sun Jun 06 2004, Hits: 1413, Reviews: 0 Write review