Illumina HiSeq 2000 paired end sequencing
Illumina WGS paired-end sequencing of additional Iberian lynx Pavon.
The standard Illumina protocol with minor modifications was followed for the creation of short-insert paired-end libraries (Illumina Inc., Cat. # PE-930-1001). In brief, 2.0 µg of genomic DNA was sheared on a Covaris E220 in order to reach the fragment size of ~500bp. The fragmented DNA was end-repaired, adenylated and ligated to Illumina specific paired-end adaptors. To obtain a library of very precise insert size (500bp) with the size deviation of +/- 25bp, the DNA with adaptor-modified ends was size selected and purified using the E-gel agarose electrophoresis system (Invitrogen). After the size selection the library was PCR amplified using 10 PCR cycles.