Improved DNA loading buffers with increased DNA detection sensitivity in both agarose-gel and polyacrylamide-gel electrophoreses.

Haroon, M. and Li, X.-Q. (2008). "Improved DNA loading buffers with increased DNA detection sensitivity in both agarose-gel and polyacrylamide-gel electrophoreses.", Journal of Huazhong Agricultural Univerity / Huazhong nongye daxue xuebao, 27(9), pp. 32-36.

Abstract

Loading buffer is a basic component in DNA electrophoresis protocols. Most buffers are only suitable for either agarose or polyacrylamide gels. Although existing loading buffers can meet the need for general electrophoresis, any improvement of the gel resolution by the loading buffer can be important when high resolution, detection sensitivity, and reproducibility are required such as in quantitative DNA analysis and multiplex PCR DNA fingerprinting. In this study, we tested five commercially available DNA loading buffers and five newly designed ones six times using both agarose gels and polyacrylamide gels (both were native). The shape,sharpness,and brightness of DNA bands were considerably influenced by the loading buffers. Among the loading buffers tested, the best one for agarose gels was LB6 (its 6-fold concentrate contains 17 % Ficoll-400,50 mmol/L Trisebase, 50 mmol/L pH 8. 0 EDTA, 0. 05 % bromophenol blue, & 0. 1 % xylene cyanol FF). For polyacrylamide gels, the best loading buffer was LBlO (its 6-fold concentrate contains 40% sucroge, 50 mmol/L pH 8. 0 EDTA, 0. 1% bromophenol blue, and 0. 1 % xylene cyan01 FF). When used the same loading buffer for both agarose gels and polyacrylamide gels,LBlO was the best. Without using the delicate silver staining of gels, DNA size ladders and p~l~merasechain-reaction-amplifiedD NA showed high quality bands in photography after simple ethidium bromide staining or GelRed staining when loaded with LB10 in polyacrylamide gel electrophoresis.