Next Generation Sequencing Method for Illumina TruSeq DNA Sample Preparation Protocol on the Hamilton STAR

Next Generation Sequencing (NGS) has accelerated the sequencing of genomes and this technology has a major role to play in the biomedical research. The simplicity of NGS library preparation, the ability to make these libraries from tiny input amounts of DNA, the elimination of a bacterial cloning intermediate and the development of data-analysis approaches to support the increasing size of the sequence datasets have been invaluable in the advancement of NGS. This application note describes the automation of the Illumina TruSeq DNA sample preparation on the Hamilton STAR . The data presented includes PicoGreen assay and Sybr E-Gel results for comparison of library construction using automated and manual methods, Bioanalyzer and KAPA assay results and data from Illumina HiSeq. Automation allowed processing 96 samples simultaneously in less than 4 hrs. Sample recovery yields higher by 50% were obtained with automated compared to manual method. The sample preparation provided average 80% reads on target from Illumina HiSeq.