GSM7574961: Anther, Lyc-B1; Lycium barbarum; RNA-Seq
Peel off the anthers and place quickly on ice,RNA was havested using Trial reagent,Illumina HiSeq/MiSeq sequencing. After the samples were eligible for detection, eukaryote mRNA was enriched with magnetic beads bearing Oligo (DT)(in the case of prokaryotes, the mRNA was enriched by kit removal of rRNA) . Fragmentation buffer was then added to break up the mRNA into short fragments and use the mRNA as a template to synthesize one-stranded cDNA using six-base random primers (random hexamers) . Then buffer, dNTPs and DNA polymerase I and RNase H were added to synthesize two-stranded cDNA, the double-stranded cDNA was purified with AMPure XP beads. The purified double-stranded cDNA was repaired, a-tailed and connected with sequencing linker, then the fragment size was selected with AMPure XP beads. The PCR products were purified with AMPure XP beads and the final library was obtained. After the library was built, Qubit2.0 was used for initial quantification, diluting the library to 1.5 ng/ul, and then Agilent 2100 was used for insert size detection of the library