Illumina NovaSeq 6000 paired-end Sequencing of KAS22054415 (KAE22249810)
Construction of reference genome of 'Baekdansim' and development of cold resistance markers
Truseq Nano DNA libraries were constructed by manufacturer's instructions. In total, 100 ng or 200 ng of high molecular weight genomic DNA for small 350 bp or large 550bp insert size were sheared to yield DNA fragments using Covaris S2 system, respectively. Blunt-ended DNA fragments are generated using a combination of fill-in reactions and exonuclease activity. A single A-base was then added to the blunt ends of each strand, preparing them for ligation to the indexed adapters. Each adapter contained a single T-base overhang for ligating the adapter to the A-tailed fragmented DNA. Ligated products were amplified with reduced-bias PCR. The quality of the amplified libraries was verified by capillary electrophoresis Bioanalyzer, Agilent. After QPCR using SYBR Green PCR Master Mix Applied Biosystems, we combined libraries that index tagged in equimolar amounts in the pool. WGS sequencing is performed using an Illumina NovaSeq 6000 system following provided protocols for 2x100 sequencing.