title: N. vectensis Chicago Library 2
design: 2 Chicago libraries were prepared as described previously . For each library, ~500 ng of HMW gDNA (mean fragment length = 100 kbp) was reconstituted into chromatin in vitro and fixed with formaldehyde. Fixed chromatin was digested with DpnII, the 5 overhangs filled in with biotinylated nucleotides, and then free blunt ends were ligated. After ligation, crosslinks were reversed and the DNA purified from protein. Purified DNA was treated to remove biotin that was not internal to ligated fragments. The DNA was then sheared to ~350 bp mean fragment size and sequencing libraries were generated using NEBNext Ultra enzymes and Illumina-compatible adapters. Biotin-containing fragments were isolated using streptavidin beads before PCR enrichment of each library. The libraries were sequenced on an Illumina HiSeq 2500 (rapid run mode). The number and length of read pairs produced for each library was: 116 million, 2x101 bp for library 1; 35 million, 2x101 bp for library 2. Together, these Chicago library reads provided 125 x sequence coverage of the genome (1-100 kb pairs).