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Unlimited increase in the resolution of DNA ladders
Author(s) -
Griess Gary A.,
Rogers Eric,
Serwer Philip
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(20000301)21:5<859::aid-elps859>3.0.co;2-5
Subject(s) - pulsed field gel electrophoresis , dna , gel electrophoresis of nucleic acids , gel electrophoresis , electrophoresis , resolution (logic) , agarose gel electrophoresis , agarose , high resolution , microbiology and biotechnology , biology , chemistry , chromatography , genetics , computer science , gene , genotype , geology , remote sensing , artificial intelligence
Fractionation of DNA ladders by gel electrophoresis is limited by the progressive compressing of the long DNA end of a ladder. Improvement in the resolution of this DNA is achieved by use of the following two‐step electrophoresis. Initially, the DNA ladder is fractionated by conventional constant field agarose gel electrophoresis. Subsequently, gel electrophoresis is performed in the reverse direction by pulsing the electrical field (PFGE). A newly developed type of pulsing is used, which causes inversion of a double‐stranded DNA ladder: the distance migrated increases as the length of the DNA molecule increases. Thus, the resolution of DNA bands continues to increase during the PFGE. These two stages of electrophoresis are serially repeated. Eventually, both the short and the long DNA ends of the ladder migrate out of the gel while a selected region of the ladder undergoes progressive increase in resolution during back‐and‐forth migration. Improved resolution of DNA bands is achieved, without a known limit.