Whole Genome Sequencing of IISR 853
Quality of DNA was checked on 0.8% agarose gel (loaded 3 l) for the single intact band. The gel was run at 110 V for 30 mins. 1 l of each sample was used for determining concentration using Qubit 2.0 Fluorometer. The paired-end sequencing library was prepared using NEBNext Ultra DNA Library Prep Kit for Illumina. The library preparation process was initiated with 200ng DNA. The DNA was mechanically sheared into smaller fragments by Covaris followed by continuous step of end-repair where an A is added to the 3 ends making the DNA fragments ready for adapter ligation. Following this step, platform-specific adapters are ligated to both ends of the DNA fragments. These adapters contain sequences essential for binding dual-barcoded libraries to a flow cell for sequencing, allowing for PCR amplification of adapter-ligated fragments, and binding standard Illumina sequencing primers. To ensure maximum yields from limited amounts of starting material, a high-fidelity amplification step was performed using HiFi PCR Master Mix. The amplified library was analyzed in Bioanalyzer 2100 (Agilent Technologies) using High Sensitivity (HS) DNA chip as per manufacturer's instructions. After obtaining the Qubit concentration for the library and the mean peak size from Bioanalyser profile, library will be loaded onto illumina platform for cluster generation and sequencing.