Identifying sequence similarities between DNA molecules

An atomic force microscope (AFM) imaging technique is described to compare sequences between two different DNA molecules and precisely locate nonhomologies in DNA strands. Sequence comparisons are made by forming heteroduplexes between the two molecules and, by AFM imaging the intact molecules forme...

Full description

Saved in:
Bibliographic Details
Published in:Ultramicroscopy 2000-02, Vol.82 (1), p.237-244
Main Authors: Hoyt, Peter R, J. Doktycz, Mitchel, Modrich, Paul, Warmack, Robert J, Allison, David P
Format: Article
Language:English
Subjects:
Atomic force microscope
Atomic force microscopy
BASIC BIOLOGICAL SCIENCES
Cloning, Molecular
Deoxyribonuclease EcoRI
DNA
DNA - chemistry
DNA sequences
DNA SEQUENCING
Enzymes
Genetic engineering
Genomic research
Heteroduplex
Microscopy, Atomic Force - methods
Molecular dynamics
Mutagenesis
Mutagenesis, Insertional
Nucleic Acid Heteroduplexes - chemistry
PATTERN RECOGNITION
Plasmids - chemistry
Sequence Deletion
Sequence Homology
Sequence verification
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:An atomic force microscope (AFM) imaging technique is described to compare sequences between two different DNA molecules and precisely locate nonhomologies in DNA strands. Sequence comparisons are made by forming heteroduplexes between the two molecules and, by AFM imaging the intact molecules formed, identifying both homologous and nonhomologous regions. By forming heteroduplexes between linearized wildtype pSV- β-galactosidase plasmid (6821 bp) and a series of deletion mutants we have identified nonhomologies (deletions) as small as 22 bp and as large as 418 bp. Furthermore, by incorporating our technique for AFM-mediated restriction mapping of DNA these mutations can be positioned relative to E coRI restriction sites. These results suggest AFM can be useful in identifying molecular level similarities and differences in DNA.
ISSN:0304-3991
1879-2723
DOI:10.1016/S0304-3991(99)00153-9