RNA-Seq of Phaeodactylum tricornutum: 12 uM phytate treated
1)mRNA enrichment: Oligo dT Selection or rRNA depletion; 2)RNA fragment and reverse transcription: Fragment the RNA and reverse transcription to double-strand cDNA (dscDNA) by N6 random primer; 3)End repair, add A tailing and adaptor ligation:The synthesized cDNA was subjected to end-repair and then was 3 adenylated. Adaptors were ligated to the ends of these 3 adenylated cDNA fragments; 4)PCR amplification:The ligation products were purified and many rounds of PCR amplification were performed to enrich the purified cDNA template using PCR primer; 5)Denature and cyclization:Denature the PCR product by heat and the single strand DNA is cyclized by splint oligo and DNA ligase; 6) Sequencing on DNBSEQ MGI2000 platform.