GSM3659352: Termination period of calcification in low strength group [LST-1]; Gallus gallus; RNA-Seq
Total RNA was isolated using the Trizol Reagent, after which the concentration, quality and integrity were determined using a NanoDrop spectrophotometer. Sequencing libraries were generated using the TruSeq RNA Sample Preparation Kit(Illumina, USA). First strand cDNA was synthesized using random oligonucleotides and SuperScript II. Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H. Remaining overhangs were converted into blunt ends via exonuclease/polymerase activities and the enzymes were removed. After adenylation of the 3′ ends of the DNA fragments, Illumina PE adapter oligonucleotides were ligated to prepare for hybridization. To select cDNA fragments of the preferred 200 bp in length, the library fragments were purified using the AMPure XP system (Beckman Coulter,Beverly, CA, USA). DNA fragments with ligated adaptor molecules on both ends were selectively enriched using Illumina PCR Primer Cocktail in a 15 cycle PCR reaction. Products were purified (AMPure XP system) and quantified using the Agilent high sensitivity DNA assay on a Bioanalyzer 2100 system (Agilent).
GEO Accession
GSM3659352