RNA-Seq of ZM4: substr 3.6M at pH 6
rRNA was removed with kit after total  RNA was collected from prokaryocyte. Fragmentation buffer was added for  interrupting mRNA to short fragments. Taking these short fragments as  templates, Random hexamer-primer were used to synthesize the first-strand cDNA.  The second-strand cDNA was synthesized using buffer, dATPs,dGTPs,dCTPs,dUTPs,  RNase H and DNA polymerase I respectively after removing dNTPs. Short  fragments were purified with QiaQuick PCR extraction kit and resolved with EB  buffer for end reparation and adding poly(A). After that, the short fragments  were connected with sequencing adapters. Then, the UNG enzyme was used to  degrade the second-strand cDNA, and the product was purified by MiniElute PCR  Purification Kit before PCR amplification. At last, the library could be  sequencing using filter.