PMID: 9547265Jun 20, 1998Paper

Short unligated sticky ends enable the observation of circularised DNA by atomic force and electron microscopies

Nucleic Acids Research
B Revet, A Fourcade

Abstract

A comparative study of the stabilisation of DNA sticky ends by divalent cations was carried out by atomic force microscopy (AFM), electron microscopy and agarose gel electrophoresis. At room temperature, molecules bearing such extremities are immediately oligomerised or circularised by addition of Mg2+or Ca2+. This phenomenon, more clearly detected by AFM, requires the presence of uranyl salt, which stabilises the structures induced by Mg2+or Ca2+. DNA fragments were obtained by restriction enzymes producing sticky ends of 2 or 4 nucleotides (nt) in length with different guanine plus cytosine (GC) contents. The stability of the pairing is high when ends of 4 nt display a 100% GC-content. In that case, 95% of DNA fragments are maintained circular by the divalent cations, although 2 nt GC-sticky ends are sufficient for a stable pairing. DNA fragments with one blunt end and the other sticky appear as dimers in the presence of Mg2+. Dimerisation was analysed by varying the lengths and concentrations of DNA fragments, the base composition of the sticky ends, and also the temperature. Our observation provides a new powerful tool for construction of inverted dimers, and circularisation, ligation analysis or short bases sequence intera...Continue Reading

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Citations

Sep 17, 2013·Analytical Biochemistry·Adam J BrownDavid C James
Mar 30, 2007·Nucleic Acids Research·Loïc HamonOlivier Piétrement
Mar 23, 2013·Acta Crystallographica. Section D, Biological Crystallography·Yann-Vaï Le BihanMarie-Hélène Le Du
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Jun 15, 2006·The Journal of Physical Chemistry. B·Yonghai SongTao Yang

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