FORMAMIDE MODIFIED POLYACRYLAMIDE GELS FOR DNA SEQUENCING BY CAPILLARY GEL-ELECTROPHORESIS

被引:40
作者
ROCHELEAU, MJ [1 ]
GREY, RJ [1 ]
DA, YC [1 ]
HARKE, HR [1 ]
DOVICHI, NJ [1 ]
机构
[1] UNIV ALBERTA,DEPT CHEM,EDMONTON T6G 2G2,ALBERTA,CANADA
关键词
D O I
10.1002/elps.11501301102
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:484 / 486
页数:3
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