Molecular identification of heparin from pigs by allele-specific PCR (AS-PCR) and amplification refractory mutation system (ARMS).
- Author:
Zhi-Yong YU
1
;
Ge DING
;
Xiao-Yu DING
;
Bi-Hai CHU
;
Liang QIAN
;
Sun GU
Author Information
1. College of Life Sciences, Nanjing Normal University, Nanjing 210046, China.
- Publication Type:Journal Article
- MeSH:
Alleles;
Animals;
Base Sequence;
DNA Mutational Analysis;
methods;
DNA Primers;
DNA, Mitochondrial;
genetics;
Drug Contamination;
prevention & control;
Heparin;
analysis;
genetics;
Horses;
genetics;
Molecular Sequence Data;
Phylogeny;
Polymerase Chain Reaction;
methods;
Quality Control;
Ruminants;
genetics;
Sequence Alignment;
Sequence Analysis, DNA;
Sus scrofa;
genetics
- From:
Acta Pharmaceutica Sinica
2008;43(5):535-541
- CountryChina
- Language:Chinese
-
Abstract:
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.
