Formamide modified polyacrylamide gels for DNA sequencing by capillary gel electrophoresis.
Compressions are occasionally found during the separation of DNA sequencing fragments, particularly in G/C-rich regions and in gels operated at room temperature. Addition of at least 10% formamide to urea/polyacrylamide sequencing gels improves the denaturing capacity of the gel, minimizing compressions. Addition of 20% or more formamide decreases the separation rate, theoretical plate count, and resolution for normally migrating fragments. An optimum concentration of 10% formamide improves resolution of compressed regions without degrading the other characteristics of the gel. Operation of gels at room temperature simplifies the engineering associated with automated sequencers based on capillary gel electrophoresis.
Rocheleau
M J
MJ
Department of Chemistry, University of Alberta, Edmonton, Canada.
Grey
R J
RJ
Chen
D Y
DY
Harke
H R
HR
Dovichi
N J
NJ
D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005559:Formamides; D005782:Gels / Q000737:chemistry*; D009691:Nucleic Acid Denaturation; D013696:Temperature
D005559:Formamides; D005782:Gels; C031066:formamide; D004247:DNA