Full-length cDNA library construction for leaves of Vicia unijuga with SMART technique

Full-length cDNA Library for leaves of Vicia unijuga was constructed with SMARTTM cDNA library construction kit after total RNA of V.unijuga leaves was extracted by Trizol reagent. The primary library has titer of 2.3×106 pfu·mL-1, while the amplified library has a high titer of 6.2×109 pfu·mL-1 and 92.7％ recombinant rate. Sixteen plaques were randomly selected from the amplified library and amplified by PCR. The results showed that the inserted fragment length ranged from 500 to 2 500 bp. The 3′end full-length cDNA of CAD, a Vicia unijuga gene related with lignin synthesis which has been cloned by RACE, was amplified in this cDNA library using PCR, which indicated that the cDNA library for leaves of V.unijuga had been successfully constructed using SMART technique.