P06115M1
Genomic characterization of Przewalski’s and Mongolian domestic horses by whole genome re-sequencing
DNA was isolated using a QIAmp DNA Mini Kit (Qiagen) or PaxGene DNA Prep Kit (Qaigen) according to the manufacturer’s protocol. DNA was applied to library preparation satisfying QC guidelines; concentration over 50 ng/microlitter, OD 260/280 ratio over 1.6, OD 260/230 ratio over 1.6. The gDNA was sheared using Covaris S series (Covaris, MS, USA). The fragment of the sheared DNA was end-repaired, A-tailed, and PE-adapted using a Library Preparation Kit (Illumina, CA, USA). After ligation of the adapters, the samples were purified and dissolved in 30 microlitter of elution buffer, after which 1microlitter of the mixture was used as a template for 12 cycles of PCR amplification. The PCR product was gel-purified using a QIAquick Gel Extraction Kit (Qiagen, Hilden, Germanay). Library quality and concentration was determined using an Agilent BioAnalyzer 2100 (Agilent, CA, USA), and library quantification was against PhiX control library using qPCR methods on a LightCycler 480 (Roche, IN, USA) according to Illumina’s library quantification protocol. Sequencing was performed using a 100-bp PE strategy and HiSeq Sequencing Kit on an Illumina HiSeq 2000 according to the manufacturer's instructions. A base-calling pipeline (Sequencing Control Software, SCS; Illumina) was used to process the raw fluorescent images and call sequences.
P06115M1
SEOUL NATIONAL UNIVERSITY
P06115M1