Screening and cloning of RAPD marker interrelated to cold tolerance in common carp

RAPD-PCR was performed with 200 random primers on DNA samples of Heilongjiang wild carp(Cyprinus carpio haematopterus Temminck et Schlegel),frigid-resistance strain of Red purse carp(Cyprinus carpio var.wananensis),Red purse carp(Cyprinus carpio var.wananensis),Boshi carp(Cyprinus pellegnini Tchang),cross F_(2) of frigid-resistance strain of Red purse and Boshi carp,respectively.RAG20 was identified to be a molecular marker which associated with cold tolerance traits of common carp(cyprinus carpio).Examination to F_(2) segregated individuals verified that this molecular marker was reliable.Up to now,10 molecular markers associated with cold tolerance of common carp were obtained,and the marker 5N1451c interrelated with cold tolerance Cyprinus (Cyprinus) pellegrini Tchang has been mapped on linkage 5.It further demonstrates that cold tolerance trait is a quantitative trait loci(QTL),which is controlled by small multi-genes.In addition,specific fragments from four primers above were extracted from the agarose and purified respectively.The purified products were then ligated to pMD 18-T vector which were transformed to competent E.coli DH 5α for sequencing.The sequencing of three specific fragmens has been finished.The length of the amplified fragments was 1?021?bp,422?bp,1?043?bp respectively.The parpose of this study is to redesign primers upon the sequences and convert RAPD markers into the stable SCAR markers finally.