GSM2759986: Rod-like3 [degradome sequencing]; Brassica napus; RNA-Seq
Total RNA was extracted using the TRIzol reagent (Invitrogen, CA, USA) according to the manufacturer’s instructions. The quantity and purity were assayed with an Agilent 2100 Bioanalyzer (AgilentTechnologies, Palo Alto, CA) to ensure sample reliability. Each RNA sample was generated from the shoot apexes of 6 different plants. For degradome sequencing, approximately 200 µg of total RNA were used for the purification of poly adenylated RNA molecules using the Oligotex mRMA kit (Qiagen). Then, using T4 RNA ligase (Ambion), the single-stranded 5′ RNA adaptor was ligated to the cleaved products that had a 5′-monophosphate end. The ligated products were purified and subjected to 5 PCR cycles for reverse transcription reactions, digested with MmeI, and ligated to a 3′ double-stranded DNA adaptor. Finally, the samples were subjected to another 20 cycles of PCR amplification, and gel-purified PCR products were subjected to sequencing
GEO Accession
GSM2759986