protein based standards for DNA/CE

Beth Stewart stewart at camis.stanford.edu
Tue May 23 17:41:40 EST 1995


Hi, we are currently working on a capillary electrophoresis system.  We
want to size PCR fragments, and are using thiazole orange (TO) to detect
the DNA (an interculating agent similar to EtBr)  We need a size standard
(with 3 peaks spanning the size range of interest) which we can run in the
same capillary as the unknown DNA fragments (which will run from 80 to 400
bp) but will not interact with the thiazole orange since we do not want it
to mask any sample peaks.  We were thinking that maybe we could put some
other kind of standard in the capillary which would be excitable with a 488
laser but would emit a good distance from TO (520), such as a labelled
protein standard, or plastic, or anything.  We run in a nondenaturing
system, 0.5X TBE iwth 0.5% hydroxymethylcellulose.  Any ideas would be
appreciated.  Thanks, Beth



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