[Molecular identification of heparin from pigs by allele-specific PCR (AS-PCR) and amplification refractory mutation system (ARMS)].
The aim of this study is to develop a convenient and effective method for the identification of heparin from pigs (include Sus scrofa domestica Brisson and Sus scrofa riukiuanus). Based on sequences of D-loop region of pigs and the other animals, two pairs of highly specific primers were designed for distinguishing heparin of pigs from other animals. The primers were employed to amplify D-loop region of DNA templates extracted from pig and seven other animal species that amounted to 49 samples. AS-PCR (allele-specific PCR) and ARMS (amplification refractory mutation system) were all suitable for fast identification of heparin from pig with anneal temperature at 54-56 degrees C in AS-PCR and with wider anneal temperature in ARMS,at 52-58 degrees C. An about 170 bp DNA fragments were amplified from separately pigs and whereas no DNA fragment was amplified from other samples under the same reaction condition.
Yu
Zhi-Yong
ZY
College of Life Sciences, Nanjing Normal University, Nanjing 210046, China.
Ding
Ge
G
Ding
Xiao-Yu
XY
Chu
Bi-Hai
BH
Qian
Liang
L
Gu
Sun
S
D000483:Alleles; D000818:Animals; D001483:Base Sequence; D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000235:genetics*; D004340:Drug Contamination / Q000517:prevention & control; D006493:Heparin / Q000032:analysis* / Q000235:genetics; D006736:Horses / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D011786:Quality Control; D012418:Ruminants / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D034421:Sus scrofa / Q000235:genetics*
D017931:DNA Primers; D004272:DNA, Mitochondrial; D006493:Heparin