GSM3261606: MC1 [degradome]; Oryza sativa; RNA-Seq
Total RNA was extracted using Trizol reagent (Invitrogen, CA, USA) following the manufacturer's procedure.  The total RNA quantity and purity were analysis of Bioanalyzer 2100 and RNA 6000 Nano LabChip Kit  (Agilent, CA, USA) with RIN number >7.0. Approximately 20 ug of total RNA were used to prepare  Degradome library. The method differed considerably from past efforts (Addo-Quaye et al., 2008, 2009b) and  followed as (Ma et al., 2010) with some modification. (1) Approximately 150 ng of poly(A)+ RNA was used as  input RNA and annealing with Biotinylated Random Primers. (2) Strapavidin capture of RNA fragments  through Biotinylated Random Primers. (3)5' adaptor ligation to to only those RNAs containing 5'-  monophosphates. (4) Reverse transcription and PCR (5) Libraries were sequenced using the 5' adapter only,  resulting in the sequencing of the first 36 nucleotides of the inserts that represented the 5' ends of the original  RNAs. And then we performed the single-end sequencing (36 bp) on an Illumina Hiseq2500 at the LC-BIO  (Hangzhou, China) following the vendor's recommended protocol. RNA libraries were prepared for sequencing using standard Illumina protocols
GEO Accession
GSM3261606