title: WGS of Nicotiana attenuata MAGIC RIL: L2_153
design: Seedlings were grown in Jiffies and Tekus until large enough for harvest (ca. 23 weeks) with a day/night cycle of 16h (26-28 C)/ 8h (22-24 C) in glasshouse at Max Planck Institute for Chemical Ecology, Jena, Germany. Frozen tissue was shipped to Novogene and gDNA extraction and sequencing was perfomed at Novogene HK. A total amount of 1.0g DNA per sample was used as input material for the DNA sample preparations. Sequencing libraries were then generated using NEBNext DNA Library Prep Kit following manufacturer's recommendations and indices were added to each sample. The genomic DNA is randomly fragmented to a size of 350bp by shearing, then DNA fragments were end polished, A-tailed, and ligated with the NEBNext adapter for Illumina sequencing, and further PCR enriched by P5 and indexed P7 oligos. The PCR products were purified (AMPure XP system) and resulted libraries were analyzed for size distribution by Agilent 2100 Bioanalyzer and quantified using real-time PCR. The qualified libraries are fed into Illumina Hiseq X after pooling according to its effective concentration and expected data volume.