[Development of a rapid mismatch PCR method using tetra-primer ARMS for detection of BMPR-IB gene mutation in sheep].
Tetra-primer ARMS PCR is a rapid, simple and efficient method for the detection of single nucleotide polymorphism (SNP). Bone morphogenetic protein receptor IB (BMPR-IB) gene is a major effect gene that controls prolific trait in Booroola Merino sheep. The aim of this study was to establish a tetra-primer ARMS PCR method for the detection of BMPR-IB genotype. Specific primers around the mutation locus (A746G) were designed according to the principle of tetra-primer amplification refractory mutation system PCR. Furthermore, a pair of primers were designed to amplify the DNA fragment, which contain the mutation locus as positive control, and an individual genotype was directly detected by one round of PCR amplification. The detection efficiency of sheep BMPR-IB genotyping was greatly improved by use of this method.
Guan
Feng
F
Research Institute of Animal Breeding & Reproduction of NanJing Agric Univ, NanJing 210095, China. guanfengheyh@yahoo.com.cn
Yang
Li-Guo
LG
An
Jun-Tao
JT
Liu
Shou-Ren
SR
Shi
Guo-Qing
GQ
D000818:Animals; D052005:Bone Morphogenetic Protein Receptors, Type I / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics*; D005838:Genotype; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results; D034561:Sheep, Domestic / Q000235:genetics*
D017931:DNA Primers; D052005:Bone Morphogenetic Protein Receptors, Type I