High efficiency hydrodynamic DNA fragmentation in a bubbling system

Lanhui Li, Mingliang Jin, Chenglong Sun, Xiaoxue Wang, Shuting Xie, Guofu Zhou, Albert Van Den Berg, Jan C.T. Eijkel, Lingling Shui*

*Corresponding author for this work

    Research output: Contribution to journalArticleAcademicpeer-review

    3 Citations (Scopus)
    74 Downloads (Pure)

    Abstract

    DNA fragmentation down to a precise fragment size is important for biomedical applications, disease determination, gene therapy and shotgun sequencing. In this work, a cheap, easy to operate and high efficiency DNA fragmentation method is demonstrated based on hydrodynamic shearing in a bubbling system. We expect that hydrodynamic forces generated during the bubbling process shear the DNA molecules, extending and breaking them at the points where shearing forces are larger than the strength of the phosphate backbone. Factors of applied pressure, bubbling time and temperature have been investigated. Genomic DNA could be fragmented down to controllable 1-10 Kbp fragment lengths with a yield of 75.30-91.60%. We demonstrate that the ends of the genomic DNAs generated from hydrodynamic shearing can be ligated by T4 ligase and the fragmented DNAs can be used as templates for polymerase chain reaction. Therefore, in the bubbling system, DNAs could be hydrodynamically sheared to achieve smaller pieces in dsDNAs available for further processes. It could potentially serve as a DNA sample pretreatment technique in the future.

    Original languageEnglish
    Article number40745
    JournalScientific reports
    Volume7
    DOIs
    Publication statusPublished - 18 Jan 2017

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  • Cite this

    Li, L., Jin, M., Sun, C., Wang, X., Xie, S., Zhou, G., ... Shui, L. (2017). High efficiency hydrodynamic DNA fragmentation in a bubbling system. Scientific reports, 7, [40745]. https://doi.org/10.1038/srep40745