RNA-Seq of Fusarium solani: sterile water treatment
Total RNA was extracted by TRIzol method. RNA purity and integrity were detected by RNA-specific agarose electrophoresis and Agilent 2100 Bioanalyzer to ensure that the quality of F.solani mycelium RNA met the sequencing requirements. The mRNA in total RNA was purified with a unique polyA structure. The mRNA with polyA structure in total RNA was enriched by Oligo ( dT ) magnetic beads, and the RNA was disrupted to a fragment of about 300 bp by ion disruption. Under the premise of RNA as a template, the first strand of cDNA was synthesized with 6-base random primers and reverse transcriptase, and the second strand of cDNA was synthesized with the first strand of cDNA as a template. After the library construction was completed, the fragments were enriched by PCR amplification, and the library size was about 450 bp. Then, the total concentration of the library and the effective concentration of the library were detected by Agilent 2100 Bioanalyzer. Then, according to the experimental requirements, the libraries containing different Index sequences were mixed in proportion. The mixed library was uniformly diluted to 2 nM, and the single-strand library was formed by alkali denaturation. Then sequenced