GSM5352615: Col-0 rep2; Arabidopsis thaliana; RNA-Seq
10-day-old seedlings were frozen with liquid nitrogen, and the total RNA was extracted with GeneJET™ Plant RNA Purification Kit After RNA sample quality control, high-quality RNA was obtained and used for library construction. The pipeline was consisted of five main steps:  (1) Isolated mRNA from total RNA using the oligo(dT) magnetic beads;  (2) Mixed with the fragmentation buffer and fragment the mRNA randomly;  (3) Using mRNA as the template, six-base random primers (random hexamers) were used to synthesize the first strand cDNA. Then PCR buffer, dNTPs, RNase H and DNA polymerase I were added to do second strand cDNA synthesis. AMPure XP beads are used to purify cDNAs;  (4) Purified double-stranded cDNAs were resolved with EB buffer for end reparation and single nucleotide A addition. Then AMPure XP beads were used in size selection;  (5) Finally, cDNA sequencing library was obtained by PCR amplification.