HiSeq reads from N. pusilla gDNA
The libraries were prepared using KAPA HyperPlus Kits (Roche) following the manufacturer's user guide. The initial concentration of DNA was evaluated using the Qubit dsDNA HS Assay Kit (Life Technologies). 25 ng DNA was used to prepare the libraries. Protocol starts with enzymatic fragmentation to produce dsDNA fragments followed by end repair and A-tailing to produce end-repaired, 5'-phosphorylated, 3'-dA-tailed dsDNA fragments. In adapter ligation step, dsDNA adapters are ligated to 3'-dA-tailed molecules. Final step is library amplification, which employs high fidelity, low-bias PCR to amplify library fragments carrying appropriate adapter sequences on both ends. Following the library preparation, the final concentration of the libraries (Table 1) were measured using the Qubit dsDNA HS Assay Kit (Life Technologies), and the average library size (Table 1) was determined using the Agilent 2100 Bioanalyzer (Agilent Technologies). The libraries were pooled and diluted (to 9.0pM) and sequenced paired end for 500 cycles using the HiSeq system (Illumina).