Year: 2000
Type: article
Abstract: migrate much more slowly than the ds -DNA, and a minute difference in migration distance caused by the conformation change of single strands is visualized by silver staining.Because single strands migrate slower, the area visualized is in the higher molecular weight range of gel electrophoresis.This requires longer electrophoresis times, frequently until BPB and XC run off the gel.We screened the commonly used, less expensive dyes of smaller negative charge and larger molecular weight, looking for a migration indicator useful for long electrophoresis times and visually detectable without fluorescence.Trypan blue, commonly used for vital staining, was discovered to migrate much more slowly than XC and to generate two bands visible in ordinary TBE buffers.In Figure 1, the migration distance of this dye is compared to that of XC and BPB (both from Katayama Chemicals, Nagoya, Japan) in polyacrylamide gels of different acrylamide concentration with and without glycerol.We usually put this dye in a blank gel as a migration indicator to approximate when to stop the electrophoresis used to detect polymorphism in genomic DNA (Figure 2).In our experience, this dye does not seem to harm the electrophoresis pattern of undigested and digested genomic DNAs.
Source: BioTechniques
DOI: 10.2144/00291bm07
Authors: J. Kulinski, D. Besack, C.A. Oleykowski, A.K. Godwin, A.T. Yeung
Institution: Fox Chase Cancer Center
Language: English
Subfield: Cancer Research
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,415
OpenAlex ID: W35995934