GSM2992195: f5-i/j-2; Oryza sativa; RNA-Seq
Total RNA was extracted from each sample using TRIzon Reagent (CWBIO, Beijing, China) following manufacturers' instruction. The total RNA samples are first treated with DNase I to eliminate any possible genome DNA contamination. The mRNA was isolated from 6 ug of total RNA using oligo(dT) magnetic beads. To reduce the amount of RNA secondary structure, and 5' bias of priming reverse transcription with poly(dT) primer, the mRNA is fragmented into short fragment before random-primed cDNA synthesis (Wilhelm, Marguerat et al. 2010). After double strand cDNAs were generated, the cDNA end repair and 3'-end single nucleotide A addition were performed. Then special Illumina sequencing adapters were ligated onto the cDNA pools. After the fragments were enriched by PCR amplification, qualification and quantification of the sample library were carried out with Agilent 2100 Bioanaylzer and ABI StepOnePlus Real-Time PCR System. Finally, the library products were ready for deep sequencing via Illumina HiSeqTM2000.
GEO Accession
GSM2992195