GSM3454814: N3: Leaves derived in culturing iris in swine wastewater; Iris tectorum; RNA-Seq
Total RNA was extracted using an RNAprep pure plant kit (TIANGEN Biotech Co., Ltd., Beijing, China) 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 BGISEQ-500 platform.
GEO
GSM3454814