Amplifying total RNA of ovules in Pinus tabulaeformis Carr. using Super SMART cDNA synthesis technology.

Using ovules of the female-sterile Pinus tabulaeformis Carr. mutant and the fertile, which has the similar genetic background, as materials, the total RNA was extracted with Plant RNA assistant kit. The first-strand of cDNA synthesis and double-strands cDNA amplification were performed by Super SMART cDNA synthesis technology. The quality of cDNA was identified by gradient PCR amplification cycles and electrophoresis. Their amplification efficiency was evaluated by the quantitative method. A total of 46.06 and 43.59 μg purified double-strands cDNA were amplified successfully from 1 μg total RNA,respectively, which had high quality and purity. The results indicate that Super SMART cDNA synthesis technology could amplify enough high quality cDNA from trace of total RNA. This method solves the problem of limited research materials and builds up the foundation of suppressed subtractive hybridization in future.