alias: Poly IC Treated 12 Hrs
title: miRNAseq data (FASTQ format) of Bubaline PBMC cultured for 12 hours: Poly IC Treated 12 Hr
design: Peripheral blood (about 15 ml) of healthy female Murrah buffaloes (n=4 per group; total six groups i.e. 3 treatment and 3 respective control groups) was immediately used for culturing with and without the TLR ligands as follows:  S.N. Treatment                                         Control  1. TLR4 Ligand (LPS)@ 100ng/ml for 6hr         No stimulation, 6hr  2. TLR3 Ligand (Poly I:C) @50µg/ml for 12hr No stimulation, 12hr  3. TLR9 Ligand (CpG ODN 2007) @ 10µg/ml for 12hr No stimulation, 12hr    The RNA fraction enriched for miRNAs was extracted from the PBMCs (isolated from the cultured blood samples (in 30ml of RPMI growth media, including 10%FBS, antibiotic mix, in 75cm2 culture flask), using the mirVana™ miRNA Isolation Kit (Life Technologies, USA).   The sRNA/miRNA samples were outsourced to Bioserve Biotechnologies (A CGI company, Hyderabad, India), for next generation sequencing.   The quality and quantity of the purified sRNAs were assessed using Agilent 2100 Bioanalyzer using sRNA LabChip kits (Agilent Technologies, USA).   The concentrations of sRNA and miRNA ranged between ~18,334–105,216 pg/µl and ~975–8666pg/µl, respectively.   Representative cDNA libraries were constructed using Ion Total RNA-Seq Kit v2 (Life Technologies).   The yield and size distribution of the amplified DNA was  assessed on an Agilent®2100 Bioanalyzer® instrument with the Agilent® DNA 1000 Kit. Each of the barcoded cDNA libraries were diluted to the same molar concentration (nM) and all the six libraries were sequenced by multiplexing using Ion-Torrent PI chip on the Ion Proton™ System, based on semiconductor high-throughput sequencing.
identifiers.namespace: Guru Angad Dev Veterinary and Animal Sciences Univ
identifiers.id: Poly IC Treated 12 Hrs