The Construction of SMART cDNA Library from Banana Fruit and the Screening of Actin2 Gene by Using PCR Method

Abstract:

In this paper, SMART （switching mechanism at 5＇ end of RNA transcript）was used in the construction of cDNA library of postharvested banana fruit. Total RNA was isolated from banana fruit after harvested for 2 days and cDNA was synthesized by using 15-25 round LD-PCR （Long-distance PCR）. Full length of amplified ds-cDNA （Double-strained complement DNA） was obtained and used in the construction of cDNA library afterward. The capacity of the library was measured to be 5×10⁶ Pfu ml^-1, with 93% recombinant. A cDNA was amplified by using PCR with this library diluted in 96-well-plate as templates. The sequence of this cDNA shows 83% similarity to A ctin2 gene in the NCBI database （Accession No. 692696）.