title: Illumina HiSeq 2500 sequencing; RNA-seq of ppp1r13l knockdown in neonatal mice cardiomyocytes and lipopolysaccharide stimulation kinetics transcriptome
design: RNA-seq of ppp1r13l knockdown in neonatal mice cardiomyocytes and lipopolysaccharide stimulation kinetics transcriptome
library_construction_protocol: Cardiomyocytes were isolated from neonatal mice and seeded on gelatin.  In particular, Mouse neonatal cardiomyocytes were isolated using Worthington Rat neonatal isolation kit. Briefly, hearts were isolated from 1‐ to 4‐day‐old BALB/c mice, minced, and incubated overnight at 4°C. Trypsin inhibitor was added, and heart pieces were incubated for 35 min in the presence of collagenase at 37°C. Cells were passed through 100‐μm cell strainer, fibroblasts were removed by adhesion for 2 h, and non‐adhered cells were centrifuged (50 g/5 min) and seeded at concentration of 3 × 104 cells/well in gelatin‐coated 6‐well plates in a myocytes growth medium (promocell) for 5 days.  ShRNA was delivered by lentiviruses and LPS stimulation was given after 72 hours. The results are of two biological replicates. 500 ng LPS stimulation for 2 and 4 hours RNA was extracted using Norgen RNA isolation kit.  nebnext ultra rna seq kit, multiplexed
attributes.tag: Experimental Factor: time
attributes.value: 4
attributes.tag: Experimental Factor: RNA interference
attributes.value: non-targeting shrna control
attributes.tag: Experimental Factor: stimulus
attributes.value: lipopolysaccharide