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 fingerprin...

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Bibliographic Details
Published in:Biomedical chromatography 2012-03, Vol.26 (3), p.393-399
Main Authors: Wang, Rong, Xie, Hua, Xu, Yue-bing, Jia, Zheng-ping, Meng, Xian-dong, Zhang, Juan-hong, Ma, Jun, Wang, Juan, Wang, Xian-hua
Format: Article
Language:English
Subjects:
capillary electrophoresis
codon
DNA Restriction Enzymes - metabolism
DNA, Neoplasm - analysis
DNA, Neoplasm - genetics
Electrophoresis, Capillary - methods
gastric cancer
Humans
Hydrogen-Ion Concentration
Mutation
plasmid
restriction endonuclease fingerprinting
Stomach Neoplasms - genetics
Temperature
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Summary: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. Copyright © 2011 John Wiley & Sons, Ltd.
ISSN:0269-3879
1099-0801
DOI:10.1002/bmc.1673