Establishment of Full-length Peanut Testa cDNA Library by Using Modified SMART

SMART is one of the most important methods for constructing a full-length cDNA library.In order to discover new genes from the peanut testa,the SMART method published previously was modified to improve its efficiency,reduce cost,simplify operation,and minimize test time.Using the optimized protocol,a full-length cDNA library for peanut testa was established.The new method produced an estimated titer of the amplified cDNA library as high as 3.84×109 cfu·mL-1,which was increased from 1.23×106 cfu·mL-1,with a recombinant rate of 93.3%.Furthermore,approximately 70% of the inserts,varying from 1.0 to 2.0 kb with an average size of about 1 100 bp,were obtained.The sequence analysis indicated that the full-length cDNA proportion was as high as 55%.It was concluded that the library was of desirable quality for full-length screening and cloning of the target genes.