Illumina HiSeq 2000 paired end sequencing
Illumina paired-end sequencing of sheared fosmids from lynx pardinus library (115.000 clones) 40 kb insert. reads matching vector are removed.
Illumina sequencing libraries were prepared from 90 fosmid clone pools. Each pool contained 1200 fosmid clones. Sample preparation for each fosmid pool was performed according to Illumina TruSeq DNA sample preparation guide (Illumina Inc., Catalog # PE-940-2001, Rev. C) with some modifications. In brief, 2.0 µg of genomic DNA was sheared on a Covaris E220 instrument. The fragment size (300-450bp) and quantity were confirmed with the Agilent 2100 Bioanalyzer 7500 chip. The fragmented DNA was size selected with AMPureXP beads and PEG concentration. The size selected DNA was end-repaired, adenylated and ligated to Illumina indexed paired-end adaptors.