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bioRxiv · 10.1101/367045

TnClone: high-throughput clonal analysis usingTn5-mediated library construction and de novo assembly

Hwang, B.·Heo, S.·Cho, N.·Bang, D.
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Abstract

A typical molecular cloning procedure requires Sanger sequencing for validation, which becomes cost-prohibitive and labour-intensive for large-scale clonal analysis of genotype-phenotype studies. Here we present a Tn5-mediated clonal analysis platform TnClone, which uses next-generation sequencing (NGS) to rapidly and cost-effectively analyze a large number of clones. We also developed a user-friendly graphical user interface and have provided general guidelines for conducting validation experiments. Using TnClone, we achieved more than 20-fold cost reduction compared with the cost incurred using conventional Sanger sequencing and detected low-frequency mutant clones (~10%) in mixed samples. We tested our programme and achieved 99.4% sensitivity. Our platform provides rapid turnaround with minimal hands-on time for secondary evaluation as NGS technology continues to evolve.

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Hwang, B., Heo, S., Cho, N., Bang, D.. 2018-07-11. TnClone: high-throughput clonal analysis usingTn5-mediated library construction and de novo assembly. https://doi.org/10.1101/367045

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