The Study Of Pepper Leaf CDNA Library Construction

The cDNA library of pepper leaves treated with UV radiation was constructed on the basis of optimized total RNA extraction mathod, The main results were as following:1) The total RNA extraction mathod was optimized with the pepper leaves treated with UV radiation, and the total RNA extracted through the optimized mathod could best meet the requirement of contructing the cDNA library.2) The cDNA was synthesized from the total RNA with the one point mutant of Moloney murine leukemia virus(MMLV) reverse transcriptase,and the cDNA was further size-fractioned by CHROMA SPIN-400 Column to remove the fraction shorter than 500bp. The ds cDNA was cloned into the X TriplEx2 vector and the resulting ligating matarial was X -phage packaged. The unamplied cDNA library was titered using E.coli XL-Blue, and it turned out that the titer of the cDNA library was 1.191 X 106,the number of the colonies is 1.112X 106,the titer of the amplied cDNA library was 1.532 X 1010,and the recombinant efficiency was over 80% through white/blue screening.