Improved DNA electrophoresis in conditions favoring polyborates and lewis acid complexation.
Spatial compression among the longer DNA fragments occurs during DNA electrophoresis in agarose and non-agarose gels when using certain ions in the conductive buffer, impairing the range of fragment sizes resolved well in a single gel. Substitutions using various polyhydroxyl anions supported the underlying phenomenon as the complexation of Lewis acids to DNA. We saw significant improvements using conditions (lithium borate 10 mM cations, pH 6.5) favoring the formation of borate polyanions and having lower conductance and Joule heating, delayed electrolyte exhaustion, faster electrophoretic run-speed, and sharper separation of DNA bands from 100 bp to 12 kb in a single run.
Singhal
Hari
H
Department of Biomedical Engineering, The Johns Hopkins University and the Sidney Kimmel Comprehensive Center at Johns Hopkins, Baltimore, Maryland, United States of America.
Ren
Yunzhao R
YR
Kern
Scott E
SE
D000143:Acids; D001881:Borates / Q000737:chemistry*; D004247:DNA / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D006863:Hydrogen-Ion Concentration
D000143:Acids; D001881:Borates; D004247:DNA