title: Illumina HiSeq 2500 paired end sequencing; RNA-seq analysis of 2 cell line models of lung disease (A549 and BEAS-2B) and primary bronchial epithelial cells in Project 3 of Open Targets 020 - Epigenomes of Cell Lines
design: RNA-seq analysis of 2 cell line models of lung disease (A549 and BEAS-2B) and primary bronchial epithelial cells in Project 3 of Open Targets 020 - Epigenomes of Cell Lines
library_name: NHBE_3_RNA_p
library_construction_protocol: Cells purchased from Lonza. They were thawed in February 2016 and cultured for approximately 12 days to reach a confluency of about 90%. Culture medium: BEBM (Lonza) plus human recombinant EGF, Hydrocortisone, Bovine Pituitary Extract, Epinephrine, Transferrin, Insulin, Triiodothyronine, and GA-1000. Qiagen RNeasy plus mini kit Total RNA from each sample was used as input for the Illumina TruSeq Stranded messenger RNA LT Sample Prep Kit (Illumina) and sequencing libraries were created according to the manufacturer’s protocol. Briefly, poly-A containing mRNA molecules were purified using poly-T oligo-attached magnetic beads. Following purification, the mRNA was fragmented and copied into first strand complementary DNA using random primers and reverse transcriptase. Second strand cDNA synthesis was then done using DNA polymerase I, RNase H and substituting dUTP for dTTP. The cDNA was ligated to adapters and enriched with PCR to create the final cDNA library. The library was pooled and sequenced
attributes.tag: Experimental Factor: cell line
attributes.value: NHBE