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- W4365449931 abstract "Plasmid construction is central to life science research, and sequence verification is arguably its costliest step. Long-read sequencing has emerged as a competitor to Sanger sequencing, with the principal benefit that whole plasmids can be sequenced in a single run. Nevertheless, the current cost of nanopore sequencing is still prohibitive for routine sequencing during plasmid construction. We develop a computational approach termed Simple Algorithm for Very Efficient Multiplexing of Oxford Nanopore Experiments for You (SAVEMONEY) that guides researchers to mix multiple plasmids and subsequently computationally de-mixes the resultant sequences. SAVEMONEY defines optimal mixtures in a pre-survey step, and following sequencing, executes a post-analysis workflow involving sequence classification, alignment, and consensus determination. By using Bayesian analysis with prior probability of expected plasmid construction error rate, high-confidence sequences can be obtained for each plasmid in the mixture. Plasmids differing by as little as two bases can be mixed for submission as a single sample for nanopore sequencing, and routine multiplexing of even six plasmids can still maintain high accuracy of consensus sequencing. SAVEMONEY should further democratize whole-plasmid sequencing by nanopore and related technologies, driving down the effective cost of whole-plasmid sequencing to lower than that of a single Sanger sequencing run." @default.
- W4365449931 created "2023-04-15" @default.
- W4365449931 creator A5033010189 @default.
- W4365449931 creator A5035414131 @default.
- W4365449931 date "2023-04-13" @default.
- W4365449931 modified "2023-10-17" @default.
- W4365449931 title "Barcode-free multiplex plasmid sequencing using Bayesian analysis and nanopore sequencing" @default.
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- W4365449931 doi "https://doi.org/10.1101/2023.04.12.536413" @default.
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