GSM3269874: PG-eAna-2: Early_Anagen Sample 2; Capra hircus; RNA-Seq
The skin sample was collected from the flanking region of each goat at same time point. Prior to sampling, the site was sheared, shaved and locally anesthetized with 2% lignocaine. Approximately 7mm diameter skin samples were harvested aseptically with a single-use biopsy punch. The quality of purified total RNA provided will be checked on the Agilent Tapestation using the RNA cartridge. Total RNA with RNA Integrity Number (RIN) value greater than or equal to 8 is considered good quality. ~4ug of total RNA will be used to prepare the RNA seq library using the TruSeq RNA Sample Prep Kits (Illumina). The libraries will be prepared as per the kit protocol. In short, poly-A containing mRNA molecules will be purified using poly-T oligo-attached magnetic beads. Following purification, the mRNA will be fragmented into small pieces using divalent cations under elevated temperature. The cleaved RNA fragments will be used to synthesize first strand cDNA using reverse transcriptase and random primers followed by second strand cDNA synthesis using DNA Polymerase I and RNase H. These cDNA fragments will then go through an end repair process, the addition of a single 'A' base, and then ligation of the adapters. The products will be purified and enriched with PCR to create the final cDNA library. Bioanalyzer plots will be used at every step to assess mRNA quality, enrichment success, fragmentation sizes, and final library sizes. Both Qubit and qPCRwill be used for measuring the quantity of the library before sequencing.
GEO Accession
GSM3269874