title: Illumina HiSeq 2500 sequencing
design: TruSeq Synthetic Long Reads (TSLR)
library_name: Quercus robur long fragments
library_construction_protocol: Starting from HMW extracted DNA samples, 15 libraries were prepared using the TruSeq Synthetic Long Reads (TSLR) technology (previously known as Moleculo), according to Illumina���s protocol. Briefly, 500 ng of gDNA were sheared into approximately 10Kb fragments using g-Tube (Covaris). After end repair, A tailing and adaptor ligation, ligated products were gel size-selected around 8-10 Kb range and then quantified by qPCR. Based on this qPCR quantification, the long insert library was diluted in order to distribute about 3 fg of the library into each well of a 384-well plate. After long range PCR, diluted products were tagmented and barcoded with 384 different barcoding PCR primers. The 384 barcoded libraries were pooled together, purified and size-selected. Each library was sequenced using 100 or 150 base-length read chemistry on a paired-end flow cell on the Illumina HiSeq2500 (Illumina). The TSLRs were generated using the BaseSpace workflow, the primary sequencing data were uploaded without any modification to the BaseSpace cloud and then processed using the standard Illumina workflow to generate long synthetic reads.