GSM9684949: WT2; Rattus norvegicus; RNA-Seq
To extract total RNA from primary ECs, the R6834 Total RNA Kit was used. Cells were lysed in TRK Lysis Buffer containing 20µl β-mercaptoethanol, followed by centrifugation to remove insoluble debris. The supernatant was mixed with an equal volume of 70% ethanol and applied to an RNA binding column. After washing, RNA was eluted in Nuclease-free Water. The RNA quality was assessed using Nanodrop, Qubit, gel electrophoresis, or Agilent 2100. For full-length transcriptome library construction, 500ng of high-quality total RNA was used for reverse transcription, low-cycle PCR amplification of full-length cDNA, followed by purification with AMPure beads.