Open AccessA Mutagenic Primer Assay for Genotyping of the CRHR1 Gene Rare Variant rs1876828 (A/G) in Asians: A Cost‐Effective SNP Typing
Author(s) -
Sharma Neeraj,
Awasthi Shally,
Phadke Shubha R
Publication year - 2016
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.21832
Subject(s) - genotyping , snp genotyping , genetics , primer (cosmetics) , biology , allele , polymerase chain reaction , molecular inversion probe , restriction fragment length polymorphism , snp , genotype , microbiology and biotechnology , genomic dna , typing , variants of pcr , gene , single nucleotide polymorphism , chemistry , organic chemistry
Background Today, the genetic and genomic research entered in a new era of high‐throughput genotyping technology. However, mutagenic polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP) is still a choice of genotyping method in molecular epidemiological research. It has been extensively used for the detection of risk alleles, if the target SNP has no natural discriminating restriction site. We undertook this study to develop a mutagenic primer assay for a CRHR1 rare gene variant: rs1876828 (A/G) and to determine their allele frequency in north Indian children. Methods The mutagenic primers were designed and assay conditions were optimized to perform mutagenic PCR‐RFLP in 550 subjects. The efficiency of assay and results were validated by sequencing. Results This study demonstrated that the mutagenic primer assay is feasible and applicable to discriminate CRHR1 gene rare variant rs1876828 (A/G) and the “frequency of allele “G” was 100% in north Indian asthmatics as well as normal subjects. Conclusion This method can be used for both large‐ and small‐scale study of complex genetic, where CRHR1 gene plays the pivotal roles.