Isolation and sequencing of active origins of DNA replication by nascent strand capture and release (NSCR).
J Biol Methods
; 2(4)2015.
Article
in En
| MEDLINE
| ID: mdl-26949711
ABSTRACT
Nascent strand capture and release (NSCR) is a method for isolation of short nascent strands to identify origins of DNA replication. The protocol provided involves isolation of total DNA, denaturation, size fractionation on a sucrose gradient, 5'-biotinylation of the appropriate size nucleic acids, binding to a streptavidin coated magnetic beads, intensive washing, and specific release of only the RNA-containing chimeric nascent strand DNA using ribonuclease I (RNase I). The method has been applied to mammalian cells derived from proliferative tissues and cell culture but could be used for any system where DNA replication is primed by a small RNA resulting in chimeric RNA-DNA molecules.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Language:
En
Journal:
J Biol Methods
Year:
2015
Type:
Article
Affiliation country:
United States