Genomic sequencing of wild-type and mutant Sulfolobus islandicus LAL14/1 strains.
WT and mutant Sulfolobus strains were grown in Tryptone-Sucrose-Vitamin-Yeast Extract (TSVY) medium. Cells were harvested by centrifugation from cultures in mid-log or stationary phase and DNA extracted by treatment of cells with SDS and EDTA in the presence of RNase. Following proteinase K digestion, DNA was purified by phenol/chloroform extraction and ethanol precipitation. Sequencing libraries were prepared by the Center for Genomics and Bioinformatics, Indiana University using the Illumina Nextera DNA Flex Library kit protocol. In brief, 200 ng of good quality genomic DNA was fragmented for each sample, after end repair and adapter ligation, the library was amplified with 6 rounds of PCR cycles. The amplified and cleaned libraries were pooled and loaded on the Illumina NextSeq500 sequencer using 75 cycle High sequencing kit.