Dynamic molecular combing: stretching the whole human genome for high-resolution studies.
DNA in amounts representative of hundreds of eukaryotic genomes was extended on silanized surfaces by dynamic molecular combing. The precise measurement of hybridized DNA probes was achieved directly without requiring normalization. This approach was validated with the high-resolution mapping of cosmid contigs on a yeast artificial chromosome (YAC) within yeast genomic DNA. It was extended to human genomic DNA for precise measurements ranging from 7 to 150 kilobases, of gaps within a contig, and of microdeletions in the tuberous sclerosis 2 gene on patients' DNA. The simplicity, reproducibility, and precision of this approach makes it a powerful tool for a variety of genomic studies.
Michalet
X
X
Laboratoire de Biophysique de l'ADN, Département des Biotechnologies, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris Cedex 15, France.
Ekong
R
R
Fougerousse
F
F
Rousseaux
S
S
Schurra
C
C
Hornigold
N
N
van Slegtenhorst
M
M
Wolfe
J
J
Povey
S
S
Beckmann
J S
JS
Bensimon
A
A
D002154:Calpain / Q000235:genetics; D002874:Chromosome Mapping / Q000379:methods*; D018244:Chromosomes, Artificial, Yeast; D003001:Cloning, Molecular; D003360:Cosmids; D015342:DNA Probes; D016521:Electrophoresis, Gel, Pulsed-Field; D005821:Genetic Techniques; D016681:Genome, Fungal; D015894:Genome, Human; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007527:Isoenzymes / Q000235:genetics; D009124:Muscle Proteins; D009136:Muscular Dystrophies / Q000235:genetics; D009154:Mutation; D011506:Proteins / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics; D015203:Reproducibility of Results; D017384:Sequence Deletion; D012821:Silanes; D014402:Tuberous Sclerosis / Q000235:genetics; D000077004:Tuberous Sclerosis Complex 1 Protein; D000077005:Tuberous Sclerosis Complex 2 Protein; D025521:Tumor Suppressor Proteins
D015342:DNA Probes; D007527:Isoenzymes; D009124:Muscle Proteins; D011506:Proteins; D012097:Repressor Proteins; D012821:Silanes; C000624653:TSC2 protein, human; D000077004:Tuberous Sclerosis Complex 1 Protein; D000077005:Tuberous Sclerosis Complex 2 Protein; D025521:Tumor Suppressor Proteins; C105884:CAPN3 protein, human; D002154:Calpain