DEVELOPMENT OF TOOLS FOR THE PREPARATION OF GENOMIC LIBRARIES FOR NEXT-GENERATION SEQUENCING

摘要: Recent advances in DNA sequencing technologies have allowed researchers to decrease the cost and time requirements for genomic by orders of magnitude. Investments novel methods improvements existing next-generation platforms resulted much higher accuracy lower cost-per-base using a variety chemistries. However, one bottlenecks all next- generation is amount resources required template library preparation. This process typically results considerable sample loss low throughput. Two steps that are need improvement fragmentation long strands amplification fragmented genomes. To enable creation high quality libraries, I developed an automated device, based on syringe pump, random DNA. The length resulting fragments tunable single parameter ends easily repaired efficient ligation adapters. also method unbiased linear concerted activities nicking endonuclease polymerase. optimization reaction conditions markedly better performance than similar protocols demonstrated utility this amplifying phage genome. explored highly specific homing use with technique, including incorporation non-native nucleotides engineering enzyme's catalytic site. Lastly, made significant progress efforts towards nuclease's recognition residues order create enzyme modification or large genomes