title: Elucidating the shelled gastropods biodiversity in the intertidal rocky coasts of Peninsular Malaysia using high-throughput sequencing
design: A total of 100 ng gDNA as measured using Qubit high sensitivity dsDNA assay (Thermo Fisher Scientific, Waltham, MA) was fragmented to 300 bp using Bioruptor (Diagenode, Denville, NJ). The fragmented DNA was subsequently processed using the NEBUltra II Illumina Library Preparation Kit according to the manufacturers instructions. Briefly, the fragmented DNA was converted to blunt-ended DNA with 5 phosphates, and 3-hydroxyls followed by A-tailing to incorporate a single A overhang base at the 3 end. Then, Illumina adapters were ligated to the A-tailed DNA fragments followed by index PCR to incorporate sample-specific barcode index and to enrich DNA fragments with ligated adapters at both ends. The constructed library was analyzed on a Fragment Analyzer (Agilent, Santa Clara, CA) to estimate its concentration and fragment size distribution. Sequencing of the constructed library used NovaSEQ6000 (2 x 150 bp configuration).