RNA-Seq of Anther: anther at binucleate
All samples were given to Sagene Biotech Co. Ltd (Guangzhou, China) for library and sequencing, and the eukaryotic mRNA was enriched with Oligo (dT) beads. And then the fragmentation buffer was used to interrupt the mRNA into short fragments to mRNA. cDNA is synthesized by using mRNA as a template by using random primers. DNA polymerase I, RNase H, dNTP and buffer were added to synthesize Second-strand Then those were purified with QiaQuick PCR extraction kit (GeneStar Co. Ltd, Beijing, China). The purified cDNA was needed to end repaired, poly (A) added. Finally, PCR amplification was performed and the PCR product was purified with AMPure XP beads Beckman Coulter, Brea, CA, USA to obtain a final library. After the library is qualified, different libraries are pooled according to the requirements of the effective concentration and the target data volume, and then the Illumina HiSeq sequence is performed.