Establishment and Application of Multiplex Amplification System for Horse STR
This article aims to construct and valuated a novel multiplex PCR system,which consists of 21 loci and is used to identify the parentage of horses.Applying fluorescent labeling technology,HTG06,HTG07,COR022,CA425,HTG04,COR082,LEX54,COR069,AHT05,HMS03,HMS01,HMS06,HMS07,COR058,HTG10,ASB17,VHL20,HMS02,ASB02 LEX34,Amelogenin,a total of 21 equine motifs,were subjected to primer design,PCR complex amplification and CE platform sequencing to verify the sensitivity,specificity and tolerance of the system.A total of 128 actual samples were tested and compared for genetic relationship.Among them,66 samples with unknown genetic relationship were analyzed by polymorphism analysis.The complex amplification system was able to detect 128 equine samples to obtain complete STR typing,and in the actual amplification,no non-specific peaks were generated,and no specific amplification peaks appeared for different species of animals,such as bovine,sheep,pig,chicken,cat,dog and human DNA samples.0.075 ng of equine standard could still obt ain complete typing under 10 μL system.The tolerance of the four common inhibitors could reach 200μmo l/L for heme,200 μmol/L for hemoglobin,100 ng/μL for humic acid and 1.2 mmol/L for EDTA,respectively.The STR multiplex amplification system established in this study is effective and reliable,which can meet the needs of laboratory for the identification of paternity of horses.
万方数据
维普
