|Public on Jan 09, 2018
|expandable arterial endothelial precursors (eAEPs)
|cell type: mature endothelial progeny of eAEP line MS42, obtained by the downregulation of MYCN and SOX17 for four days
culture medium: EGM2 (Lonza) supplemented with 10 mM HEPES
tissue of origin: cord blood
|from RLT+ cell lysates using the RNeasy Plus Micro Kit (Qiagen) with genomic DNA elimination (gDNA Eliminator Spin Columns)
The following is an excerpt from Hou et al, Sci Rep. 2015; 5:9570, the manuscript that details the protocol employed. Poly-A-tailed mRNA is isolated from total RNA using oligo-dT beads. Purified mRNA is then fragmented with heat in fragmentation buffer. First strand cDNA is then synthesized using random hexamer oligos containing partial Illumina 3′ adaptor sequence. After RNA removal, a modified oligo containing partial Illumina's 5′ adaptor is then ligated to the 5′ of the single stranded cDNA. The library is then amplified by PCR using oligos that contain full Illumina adaptor sequences and our in-house index sequences.
|Illumina HiSeq 2500
|Basecalls/demuxxing performed using Illumina bcl2fastq v1.8.4.
Reads were filtered and trimmed for adapters and quality. Expression estimates were calculated using RSEM v1.2.3 with alignments by bowtie v0.12.9
Supplementary_files_format_and_content: gzipped tab-delimited file containing RSEM's calculated Expected Counts gene expression estimates.
|Nov 16, 2017
|Last update date
|May 15, 2019
|Morgridge Institute for Research
|330 North Orchard Street
|RNA-seq analysis of human expandable arterial endothelial precursors (eAEPs), expandable mesenchymal precursors (eMPs), and controls