Premium
Improving the length‐fractionation of DNA during capillary electrophoresis
Author(s) -
Griess Gary A.,
Serwer Philip
Publication year - 2001
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200112)22:20<4320::aid-elps4320>3.0.co;2-s
Subject(s) - capillary electrophoresis , electrophoresis , fractionation , chromatography , chemistry , dna , capillary action , analytical chemistry (journal) , gel electrophoresis of nucleic acids , aqueous solution , materials science , biochemistry , composite material
The present study develops a path‐lengthening strategy for capillary electrophoresis of short double‐stranded DNA molecules, in an aqueous solution of neutral polymer (hydroxypropylmethylcellulose). Tests of the dependence of fractionations on pulse times reveal the operation of at least one mechanism in addition to increase in effective path length. Electrophoresis is performed in the following two‐stage cycles (cyclic electrophoresis): The first analysis‐stage of each cycle is a constant field (forward) capillary electrophoresis. This analysis‐stage reveals the length distribution of the shortest DNA molecules not previously analyzed. The second, enhancement‐stage of each cycle is zero‐integrated field electrophoresis (ZIFE). The enhancement‐stage improves the DNA length‐fractionation for the next DNA molecules to be analyzed. A slight reverse migration occurs in the enhancement‐stage. Increase in both peak separation and peak sharpness contribute to improvement in the length‐fractionation of DNA molecules.