CONSTRUCTION OF cDNA LIBRARY OF GERBIL′S COCHLEA / 解放军医学杂志

ABSTRACT

To construct a cDNA library of gerbil′s cochlea, the mRNA was separated from the cochlea of gerbil and the first strand cDNA was synthesized through reverse transcription by a modified oligo(dt) primer. The double strand cDNA was amplified by Ld pcR. After cDNA size fractionation , the ds cDNA was ligated in the ? TripIEx2 vector . The cDNA library was identified with special primers of prestin genes of cochlea by PCR .The results showed that the titer of library was 1.8?10 6 pfu and the percentage of recombinant clones was 80% . Prestin gene was contained in the library , the size of the fragment was 863bp . The results suggest that the established cDNA library has a high titer, recombinant percentage and large insert fragments of genes . The study lays the foundation of molecular biological study of the cochlea.