GSM1603935: WT_DPA_1; Arabidopsis thaliana; RNA-Seq
Total RNA extraction using RNeasy Plant Mini Kit (Qiagen) Multiplex RNA library construction and Illumina sequencing were carried out at the Beijing Genome Institute (BGI). The total RNA samples are first treated with DNase I. Then the mRNA is enriched by using the oligo(dT) magnetic beads. Mixed with the fragmentation buffer, the mRNA is fragmented into short fragments. Then the first strand of cDNA is synthesized by using random hexamer-primer. Buffer, dNTPs, RNase H and DNA polymerase I are added to synthesize the second strand. The double strand cDNA is purified with magnetic beads. End reparation and 3'-end single nucleotide A (adenine) addition is then performed. Finally, sequencing adaptors are ligated to the fragments. The fragments are enriched by PCR amplification. During the QC step, Agilent 2100 Bioanaylzer and ABI StepOnePlus Real-Time PCR System are used to qualify and quantify of the sample library. Approximately 20 million reads of 50 bp single-ended sequences were generated per sample using Illumina HiSeq2000.
GEO Accession
GSM1603935