title: Study on detection of mutation DNA fragment in gastric cancer by restriction endonuclease fingerprinting with capillary electrophoresis.
abstract: The DNA fragment detection focusing technique has further enhanced the sensitivity and information of DNA targets. The DNA fragment detection method was established by capillary electrophoresis with laser-induced fluorescence detection and restriction endonuclease chromatographic fingerprinting (CE-LIF-REF) in our experiment. The silica capillary column was coated with short linear polyarclarylamide (SLPA) using nongel sieving technology. The excision product of various restricted enzymes of DNA fragments was obtained by REF with the molecular biology software Primer Premier 5. The PBR322/BsuRI DNA marker was used to establish the optimization method. The markers were focused electrophoretically and detected by CE-LIF. The results demonstrate that the CE-LIF-REF with SLPA can improve separation, sensitivity and speed of analysis. This technique may be applied to analysis of the excision product of various restricted enzymes of prokaryotic plasmid (pIRES2), eukaryote plasmid (pcDNA3.1) and the PCR product of codon 248 region of gastric cancer tissue. The results suggest that this method could very sensitively separate the excision products of various restricted enzymes at a much better resolution than the traditional agarose electrophoresis.
authors.affiliation: Department of Pharmacy, Lanzhou General Hospital of PLA, No. 333 Binghe South Road, Lanzhou, Gansu, 730050, People's Republic of China. wangrong-69@163.com
authors.forename: Rong
authors.initials: R
authors.lastname: Wang
authors.forename: Hua
authors.initials: H
authors.lastname: Xie
authors.forename: Yue-Bing
authors.initials: YB
authors.lastname: Xu
authors.forename: Zheng-Ping
authors.initials: ZP
authors.lastname: Jia
authors.forename: Xian-Dong
authors.initials: XD
authors.lastname: Meng
authors.forename: Juan-Hong
authors.initials: JH
authors.lastname: Zhang
authors.forename: Jun
authors.initials: J
authors.lastname: Ma
authors.forename: Juan
authors.initials: J
authors.lastname: Wang
authors.forename: Xian-Hua
authors.initials: XH
authors.lastname: Wang
mesh_terms: D004262:DNA Restriction Enzymes / Q000378:metabolism; D004273:DNA, Neoplasm / Q000032:analysis* / Q000235:genetics; D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009154:Mutation; D013274:Stomach Neoplasms / Q000235:genetics*; D013696:Temperature
chemical_list: D004273:DNA, Neoplasm; D004262:DNA Restriction Enzymes