GSM1695168: OE6-2; Citrus unshiu; RNA-Seq
Total RNA was extracted by Trizol method The total RNA samples are first treated with DNase I to degrade any possible DNA contamination. Then the degestion products are purified by magnetic beads. After that, the mRNA is enriched by using the oligo(dT) magnetic beads (for eukaryotes). Mixed with the fragmentation buffer, the mRNA is fragmented into short fragments (about 200 bp). Then the first strand of cDNA is synthesized by using random hexamer-primed reverse transcription. 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 is then performed. After the previous step, adaptors are ligated to the ends of these fragments. Next, ligation products were selected by size and purified on TAE-agarose gel. Finally, the fragments are enriched by PCR amplification, then purified by magnetic beads and dissolved in the appropriate amount of Epstein-Barr solution.
GEO Accession
GSM1695168