4.7 Article

A tunable filter for high molecular weight DNA selection and linked-read sequencing

Journal

LAB ON A CHIP
Volume 20, Issue 1, Pages 175-184

Publisher

ROYAL SOC CHEMISTRY
DOI: 10.1039/c9lc00965e

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Funding

  1. project Gene Extractor from Region Midi-Pyrenees
  2. ANR muLAS [ANR-16-CE18-0028-01]
  3. ANR BioPulse [ANR-16-ASTR-0020]
  4. Agence Nationale de la Recherche (ANR) [ANR-16-CE18-0028, ANR-16-ASTR-0020] Funding Source: Agence Nationale de la Recherche (ANR)

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In third generation sequencing, the production of quality data requires the selection of molecules longer than similar to 20 kbp, but the size selection threshold of most purification technologies is smaller than this target. Here, we describe a technology operated in a capillary with a tunable selection threshold in the range of 3 to 40 kbp controlled by an electric field. We demonstrate that the selection cut-off is sharp, the purification yield is high, and the purification throughput is scalable. We also provide an analytical model that the actuation settings of the filter. The selection of high molecular weight genomic DNA from the melon Cucumis melo L., a diploid organism of similar to 0.45 Gbp, is then reported. Linked-read sequencing data show that the N50 phase block size, which scores the correct representation of two chromosomes, is enhanced by a factor of 2 after size selection, establishing the relevance and versatility of our technology.

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