Application of improved gel electrophoresis to the detection of genotype polymorphisms
10.3760/cma.j.issn.2095-428X.2014.15.014
- VernacularTitle:改良凝胶电泳方法在基因多态性检测中的应用
- Author:
Peige XIA
;
Qian ZHANG
;
Zanyang SHI
;
Xinru CHENG
;
Jianguo WEN
- Publication Type:Journal Article
- Keywords:
Polyacrylamide gel electrophoresis-silver staining;
Agarose gel electrophoresis-EB;
Improved gel electrophoresis;
Genotype polymorphisms
- From:
Chinese Journal of Applied Clinical Pediatrics
2014;29(15):1174-1176
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the traits of improved polyacrylamide gel electrophoresis (PAGE)-ethidium bromide(EB) staining in the detection of genotype polymorphisms.Methods The methods of PAGE-silver staining,agarose gel electrophoresis (AGE)-EB staining and improved PAGE-EB staining in polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were used to detect the μ-opioid receptor (OPRM1) A118G genotypes in case group (n =167) infants with intracranial hemorrhage (ICH),and control group(n =163) infants without ICH,to conduct a case study analysis.And the application traits of three methods were compared.Results Genotypes of OPRM1 A118G were GG (169 bp),AG (193 bp,169 bp),AA (193 bp).Both the electrophoresis methods of PAGE-silver staining and PAGE-EB could be used to detect the genotypes of OPRM1 A118G clearly in this study.There was no statistically significant difference between the resolutions of DNA fragments (P =0.884).The first method,which had 13 experiment steps,consuming 4-5 hours,involving 12 kinds of chemical reagents,and the pictures were taken with the camera,was complex,with difficult operation,more time consuming; Compared with the first method,the secondmethod was simple,which had 6 test procedures,consuming 2 hours with 8 reagents,and the pictures were taken by using an automatic gel imager.AGE-EB could not be used to detect genotypes of OPRM1 A118G.Conclusions The method of improved PAGE-EB has the advantages of fast,easy operation,and high resolution,which is worthy of wider application.
