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Nucleic Acids Research, Vol 25, Issue 8 1660-1661, Copyright © 1997 by Oxford University Press

ARTICLES

Large scale preparation of positively supercoiled DNA using the archaeal histone HMf

WA LaMarr, KM Sandman, JN Reeve and PC Dedon
Division of Toxicology, 56-787, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA.

A technique to prepare relatively large quantities (>/=100 microg) of highly positively supercoiled DNA is reported. This uses a recombinant archaeal histone (rHMfB) to introduce toroidal supercoils, and an inexpensive chicken blood extract to relax unrestrained superhelical tension. Preparation of positively supercoiled pUC19 DNA molecules, >50% of which have linking number changes ranging from+8 to+17, is demonstrated. Advantages include the high degree of positive supercoiling that can be achieved, control over the extent of supercoiling, easy production of relatively large quantities of supercoiled DNA, and low cost.
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 Proc. Natl. Acad. Sci. USAHome page
W. A. LaMarr, L. Yu, K. C. Nicolaou, and P. C. Dedon
Supercoiling affects the accessibility of glutathione to DNA-bound molecules: Positive supercoiling inhibits calicheamicin-induced DNA damage
PNAS, January 6, 1998; 95(1): 102 - 107.
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