1,721,049 research outputs found
Using archaeal histones for precise DNA fragmentation , 20: 267-271 (2007)
No full textThe fragmentation of DNA is a useful procedure for
many molecular biology procedures. However, most
methods used to fragment DNA are poorly controllable,
and cannot be used to create small fragments. We
describe a method to generate random DNA fragments of
a predictable size to be cloned in expression vectors
for the construction of display libraries. The DNA is
allowed to form complexes with archaeal histones from
Methanothermus fervidus (HMf) and the HMf/DNA core
complex is naturally protected from nuclease DNaseI
activity, giving rise to DNA fragments of 60 bp and
multiples thereof. We found that by varying the wt/wt
ratio between DNA and HMf, the concentration of DNA
and the incubation time with DNaseI, DNA fragments of
desired size can be obtained. This approach should be
applicable to the efficient fragmentation of DNA for the
construction of phage display polypeptide libraries, as
well as any other molecular biology procedures in which
small DNA fragments of defined size are required
- …
