Title
Purification and staining of intact yeast DNA chromosomes and real-time observation of their migration during gel electrophoresis
Date Issued
15 August 1997
Access level
open access
Resource Type
journal article
Author(s)
Universidad de Oregon
Publisher(s)
Portland Press Ltd
Abstract
In the past few years, fluorescence microscopy has been used successfully to characterize the motion of intermediate-size DNA molecules (50-500kbp) during steady- and pulsed-field gel electrophoresis. However, experimental difficulties had prevented the application of this technique to the direct observation of longer DNA chromosomes (1-2 Mbp). In the present study a particular procedure was followed for the purification and staining of chromosomal yeast DNA to protect it from shear forces. Also, a new highly fluorescent DNA-labelling dye, YOYO-1, was employed to improve brightness and contrast. Finally, the motion of such long DNA molecules (1-2 Mbp) was characterized under steady-field electrophoresis conditions. An accurate description of the molecular mechanisms of motion of such long molecules should provide the basis for a detailed particular procedure was followed for the purification and analysis of the mechanisms responsible for DNA trapping.
Start page
131
End page
138
Volume
326
Issue
1
Language
English
OCDE Knowledge area
Bioquímica, Biología molecular
Scopus EID
2-s2.0-0030884082
PubMed ID
Source
Biochemical Journal
ISSN of the container
02646021
Sources of information: Directorio de Producción Científica Scopus