Use of polyacrylamide gel electrophoresis to detect structural variations in kilobase-sized DNAs

✍ Scribed by Dr. Nancy C. Stellwagen

• Publisher: John Wiley and Sons
• Year: 1995
• Tongue: English
• Weight: 957 KB
• Volume: 16
• Category: Article
• ISSN: 0173-0835
• DOI: 10.1002/elps.11501601112

✦ Synopsis

Use of polyacrylamide gel electrophoresis to detect structural variations in kilobase-sized DNAs

University of Iowa, Iowa City, IA

The electrophoresis of linear, kilobase-sized DNA molecules with permuted sequences has been studied in polyacrylamide and agarose gels. Plasmid pBR322, bacteriophage QX174, and the SV40 minichromosome were each digested with a series of single-cut restriction enzymes. The linearized, permuted isomers of all three DNAs exhibit different mobilities in large-pore polyacrylamide gels, suggesting that all three DNAs contain sites of anisotropic, sequence-dependent curvature. Various experimental parameters such as acrylamide concentration, crosslinker ratio and buffer composition affect the magnitude of the observed differential mobilities. Band sharpness appears to be optimal in polyacrylamide gels containing 6.9-8.1°/oT and O.5-l0/oC. Only small mobility differences are observed for the linearized, permuted sequence isomers in agarose