A simple dialysis device for large DNA molecules

Biotechniques. 2019 Feb;66(2):93-95. doi: 10.2144/btn-2018-0133.

Abstract

The potential of genomic DNA is realized when new modalities are invented that manipulate large DNAs with minimal breakage or loss of sample. Here, we describe a polydimethylsiloxane-polycarbonate membrane device to remove small molecules from a sample while retaining large DNAs. Dialysis rates dramatically change as DNA size in kb (M) increases and DNA dimensions become comparable to pore size, and chain characteristics go from rod-like to Gaussian. Consequently, we describe empirical rates of dialysis, R, as a function of M as falling into two regimes: DNAs ≤ 1 kb show R(M) ∼e -t/τM (t = time, τM = time constant), while DNAs ≥1.65 kb slowly passage with R(M) ∼M -1.68; such partitioning potentiates single-molecule imaging.

Keywords: dialysis; large DNA; polydimethylsiloxane; small DNA; track-etched membranes.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • DNA / chemistry
  • DNA / isolation & purification*
  • Dimethylpolysiloxanes / chemistry
  • Humans
  • Membranes, Artificial*
  • Polycarboxylate Cement / chemistry
  • Single Molecule Imaging / methods*

Substances

  • Dimethylpolysiloxanes
  • Membranes, Artificial
  • Polycarboxylate Cement
  • polycarbonate
  • baysilon
  • DNA