A novel method for detecting single-nucleotide polymorphisms by using ShineRoar probes
- VernacularTitle:ShineRoar探针技术检测单核苷酸多态性
- Author:
Guang-Hua LUO
;
Lu ZHENG
;
Xiao-Ying ZHANG
;
Guo-Feng XU
;
Jiang ZHU
;
Qin-Feng MU
;
Jiang WEI
;
Lu-Jun CHEN
;
Jun ZHANG
;
Ning XU
;
- Publication Type:Journal Article
- Keywords:
Single nucleotide polymorphism;
Genotype;
Receptor,tumor necrosis factor,type Ⅱ;
Apolipoproteins
- From:
Chinese Journal of Laboratory Medicine
2000;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective The present study demonstrates a novel,simple and cost-effective method for detecting known SNP genotyping by using ShineRoar probes.Methods The SNP of target genes detected by using the ShineRoar probes and melting curve analysis.Tumor necrosis factor receptor Ⅱ (TNFR Ⅱ) and apolipoprotein M (apoM) had been employed as target genes to describe the method in details.The PCR products of TNFR Ⅱ and apoM were collected and sequenced.Results The melting temperatures (TM) were significantly different between mutated genotypes and wild-type genotype.A biallelic SNP marker (T/ G) at position 196 in exon 6 of TNFR Ⅱ gene showed two melting valleys with the appropriate TMs at (52.84?0.75)℃ and (58.38?0.61)℃,respectively.For apoM T-778C,TMs of homozygous T genotype and C genotype were (42.55?0.73)℃ and (49.19?0.57)℃,respectively.Moreover,this genotyping method was validated by the DNA sequence analyses (Kappa=1,P=0.000).Conclusion It is concluded that this novel method is simple and economical and it is suitable for a large-scale genotyping screening.
