Constructing CDNA Library Of Apple Fruits

A high quality cDNA library of M. Red stark ing fruit has been constructed. The total RNA of apple fruits has been extracted by using CTAB method. The single strand cDNA was synthesized from mRNA with MMLV reverse transcriptase, then double strand cDNA was synthesized by using PCR. Double strand cDNA size fractionation was completed by CHROMA SPIN-400 column, so the low molecular weight cDNA fragments shorter than 500 bp could be removed out. cDNA was packaged using the A. phage packaging system after being ligated to the blunted A. Trip I Ex2. The cDNA library has been constructed and amplified through transducting E.coli XL1-Blue. The constructed cDNA library has the liter of 1. 191+106 and the clone number is 1. 112 +106. While the average insert sizes of the library are 1 kb with the recombination efficiency more than 95%, and the amplified titer of the library is 1.532+1010. Therefore, a cDNA library of apple fruit has been successfully constructed.