Gap Closing/Finishing by Targeted Genomic Region Enrichment and Sequencing

Singh, Kanwar ; Froula, Jeff ; Trice, Hope ; Pennacchio, Len A. ; Chen, Feng

关键词: 99;    ALGORITHMS;    AMPLIFICATION;    DESIGN;    FUNCTIONALS Gap Closing;    Gap Finishing;    Sequencing;   

DOI  :  10.2172/985324
RP-ID  :  LBNL-3659E-Poster
PID  :  OSTI ID: 985324
Others  :  TRN: US201016%%2185

美国|英语

来源: SciTech Connect

【 摘 要 】

Gap Closing/Finishing of draft genome assemblies is a labor and cost intensive process where several rounds of repetitious amplification and sequencing are required. Here we demonstrate a high throughput procedure where custom primers flanking gaps in draft genomes are designed. Primer libraries containing up to 4,000 unique pairs in independent droplets are merged with a fragmented genomic template. From this millions of picoliter scale droplets are formed, each one being the functional equivalent of an individual PCR reaction. The PCR products are concatenated and sequenced by Illumina which is then assembled and used for gap closure. Here we present an overall experimental strategy, primer design algorithm and initial results.