Application of improved gel electrophoresis to the detection of genotype polymorphisms

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 second method 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. Key words: Polyacrylamide gel electrophoresis-silver staining; Agarose gel electrophoresis-EB; Improved gel electrophoresis; Genotype polymorphisms