EFFECT OF TOTAL PERCENT POLYACRYLAMIDE IN CAPILLARY GEL-ELECTROPHORESIS FOR DNA SEQUENCING OF SHORT FRAGMENTS - A PHENOMENOLOGICAL MODEL

被引:32
作者
HARKE, HR [1 ]
BAY, S [1 ]
ZHANG, JZ [1 ]
ROCHELEAU, MJ [1 ]
DOVICHI, NJ [1 ]
机构
[1] UNIV ALBERTA,DEPT CHEM,EDMONTON T6G 2G2,ALBERTA,CANADA
来源
JOURNAL OF CHROMATOGRAPHY | 1992年 / 608卷 / 1-2期
基金
加拿大自然科学与工程研究理事会;
关键词
D O I
10.1016/0021-9673(92)87116-P
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Polyacrylamide capillary gels were prepared with constant (5% C) cross-linker concentration and with total acrylamide concentration ranging from 2.5 to 6% T. At each acrylamide concentration, peak spacing was constant for DNA sequencing fragments ranging from 25 to 250 nucleotides in length. Peak spacing increased linearly with the total acrylamide concentration. The intercept of the retention time vs. fragment length plot was independent of %T. Ferguson plots were constructed for short DNA fragments; the polyacrylamide pore size falls in the 2.5 to 3.5 nm range for the gels studied. Theoretical plate count is independent of total acrylamide concentration; longitudinal diffusion, and not thermal gradients, limit the plate count. A phenomenological model is presented that predicts retention time, plate count, and resolution for sequencing fragments ranging in size from 25 to 250 bases and gels that range from 2.5 to 6% total acrylamide.
引用
收藏
页码:143 / 150
页数:8
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