Abstract: The cDNA libraries were constructed from Hirsutella sinensis before and after the sexual development, respectively. Total RNA KitⅠ method was used to extract total RNA. The mRNA was separated by magnetic particles, and then double stranded cDNA was synthesized by reverse transcription. The obtained cDNA was purified by proteinase K and SfiⅠdigestive enzymes and connected to the carrier pUC19, then transferred into strain DH-5α with competent cells, by electrotransformation. Afterwards, the clone number, reconstruction ratio and the size of the insert fragment of the library were measured. The results showed that library 1 contained 1.09×10⁶ recombinants, the blue and white test showed the reconstruction ratio was 92.5%, library 2 contained 1.13×10⁶ recombinants, and the blue and white test showed the reconstruction ratio was 92%. The size of reconstructed DNA fragments in two libraries ranged from 500 to 2 000 bp. It showed that RNA extracted in this experiment had good purification degree, which was enough to meet the requirements for subsequent research.

Key words: Hirsutella sinensis, sexual development, cDNA library