PRJNA526327
Homo sapiens Transcriptome or Gene expression
Full length RNA-seq - Total RNA was submitted to Pacific Biosciences (Menlo Park, CA) or Icahn School of Medicine at Mount Sinai (New York, NY). The integrity and purity of total RNA were assessed using Agilent Bioanalyzer and OD260/280 prior to submission. Full length cDNA synthesis was done from polyA RNA using Clontech SMARTer PCR cDNA synthesis kit (Clontech Laboratories; (23)). Libraries were prepared after size selection of cDNAs into bins that contain 1-2 kb, 2-3 kb and >3 kb cDNAs by the BluePippin size selection protocol. These fractions were converted to single molecule real time (SMRT) libraries followed by SMRT sequencing. 12 SMRT cells were used to sequence the lineage-negative population (5 cells 1-2 kb, 5 cells 2-3kb, 2 cells 3-6 kb) 231,960 single molecule reads uniquely associated with non-redundant isoforms. Transcripts of Full Length Unassembled (ToFU version 1) algorithm was used to create the high quality consensus transcripts from the CCS reads.