High-resolution separation of DNA restriction fragments by capillary electrophoresis in cellulose derivative...

The performance and the efficiency of several cellulose derivatives as a molecular sieving agent for the capillary electrophoretic separation of DNA restriction fragments were investigated. All fragments up to 12,000 base pairs (bp) in the 1-kbp DNA ladder were resolved using linear polyacrylamide-coated capillaries filled with a buffer solution containing 0.5% cellulose derivative and the separation was completed within 17 min. High-concentration (0.7%) cellulose derivative solutions are effective for the complete separation of small fragments (50-1000 bp) of a HincII and a HaeIII digest of phi X174 DNA. A plate number of 0.5-1 x 10(6) plates per metre was achieved. The migration time and the resolution of DNA fragments were manipulated by varying several parameters, such as the size (viscosity) and the concentration of cellulose derivatives and the applied field strength. Some guidelines are presented for choosing these parameters, depending on the size of the DNA fragments being separated.