title: Genotyping the CRHR1 rs242939 (A>G) polymorphism by a one-step tetra primer-amplification refractory mutation system-polymerase chain reaction.
abstract: OBJECTIVE
  With the rapid advances in molecular techniques, various methods for genotyping single-nucleotide polymorphisms (SNPs) are available. Still, the search for easy, robust, and less costly techniques continues. We wished to develop a Tetra primer-amplification refractory mutation system-polymerase chain reaction (T-ARMS-PCR) based technique for the corticotrophin-releasing hormone receptor 1 (CRHR1) (rs242939) SNP for use in our research lab.
  
  
  RESULTS
  To detect SNPs in a single-step PCR, we set up two genotyping methods, T-ARMS-PCR and restriction fragment length polymorphism (RFLP). The study was performed using thirty blood samples taken from clinically defined asthmatic patients. The efficiency and effectiveness of results obtained by T-ARMS were validated by PCR-RFLP and sequencing. This study demonstrates that T-ARMS is feasible and applicable to discriminate a wild-type allele from the respective mutant allele in one step.
  
  
  CONCLUSIONS
  This work is the first that presents a rapid, sensitive, and high throughput genotyping method for the CRHR1 (rs242939) polymorphism and can be used for both large- and small-scale genotyping studies.
authors.affiliation: Translational Medicine Unit, Department of Paediatrics, CSMMU, Lucknow, India.
authors.forename: Neeraj
authors.initials: N
authors.lastname: Sharma
authors.forename: Shally
authors.initials: S
authors.lastname: Awasthi
authors.forename: Subha R
authors.initials: SR
authors.lastname: Phadke
authors.forename: Sarika
authors.initials: S
authors.lastname: Gupta
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods; D017931:DNA Primers / Q000737:chemistry* / Q000235:genetics; D005260:Female; D060005:Genotyping Techniques / Q000379:methods*; D006801:Humans; D008297:Male; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide; D018019:Receptors, Corticotropin-Releasing Hormone / Q000235:genetics*; D000099052:CRF Receptor, Type 1
chemical_list: D017931:DNA Primers; D018019:Receptors, Corticotropin-Releasing Hormone; D000099052:CRF Receptor, Type 1