Animal Husbandry and Feed Science ›› 2010, Vol. 31 ›› Issue (5): 1-1.doi: 10.12160/j.issn.1672-5190.2010.05.002

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Construction and Identification of cDNA Library of Heart Tissue from Yunan Pig

YI Zhao-hui (Xinyang Vocational and Technical College,Xinyang 464000,China)   

  • Online:2010-05-20 Published:2010-05-20

Abstract: The total RNA from heart tissue of Yunan pig was extracted by isothiocyanate method in the trial.Firstly,full-length cDNAs were synthesized by SMART technique,then a pool of double-stranded cDNA was amplified by LD-PCR technology.The achived cDNA fragments was ligated into pMD18-T vector and transformed into E.coli DH5α to construct the full-length cDNA library.The library was tittered,and the rate of recombination(blue/white) was also determined.The results showed that the primary titer of the constructed cDNA library was 1.8×105 cuf/mL,the recombination rate was about 94.4% .13 transformed germs were selceted and cultured,then their plamids were extracted and identified by restriction enzyme of Hind Ⅲ and EcoRⅠ,the gel electrophoresis showed the length of most cDNA inserted in the library was about 0.3-2.0 kb.

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