Determination of the Plasmid of Protokaryon and Eukaryon by the Restriction Endonuclease Fingerprinting-Capillary Electrophoresis with Laser Induced Fluorescence Detection

Abstract

A method was developed for the determination of DNA fragments of protokaryon plasmid and eukaryon plasmid by the restriction endonuclease fingerprinting-capillary electrophoresis with laser induced fluorescence detection (REF-CE-LIF). The fused silica capillary column was coated by the nongel sieving technology of linear polyacrylamide. The molecular biology software of primer premier 5 was used to choose the cutting point of restricted enzyme. It can provide a lot of fingerprint information. The plasmid of protokaryon (pGEX-4T-1, pIRES2) and eukaryon (pcDNA3.1) were cut by the different restricted enzymes. The REF-HPCE-LIF method can very well separate the cutting product of restricted enzyme with satisfactory electropherograms of restriction endonuclease fingerprinting. The results showed that REF-CE was of higher resolution than agarose electrophoresis.