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Title: Quantitive DNA Fiber Mapping

Abstract

Several hybridization-based methods used to delineate single copy or repeated DNA sequences in larger genomic intervals take advantage of the increased resolution and sensitivity of free chromatin, i.e., chromatin released from interphase cell nuclei. Quantitative DNA fiber mapping (QDFM) differs from the majority of these methods in that it applies FISH to purified, clonal DNA molecules which have been bound with at least one end to a solid substrate. The DNA molecules are then stretched by the action of a receding meniscus at the water-air interface resulting in DNA molecules stretched homogeneously to about 2.3 kb/{micro}m. When non-isotopically, multicolor-labeled probes are hybridized to these stretched DNA fibers, their respective binding sites are visualized in the fluorescence microscope, their relative distance can be measured and converted into kilobase pairs (kb). The QDFM technique has found useful applications ranging from the detection and delineation of deletions or overlap between linked clones to the construction of high-resolution physical maps to studies of stalled DNA replication and transcription.

Authors:
; ; ; ;
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
Life Sciences Division
OSTI Identifier:
948493
Report Number(s):
LBNL-120E
TRN: US200907%%62
DOE Contract Number:  
DE-AC02-05CH11231
Resource Type:
Book
Country of Publication:
United States
Language:
English
Subject:
60; CELL NUCLEI; CHROMATIN; CONSTRUCTION; DETECTION; DNA; DNA REPLICATION; FIBERS; FLUORESCENCE; PROBES; RESOLUTION; SENSITIVITY; TRANSCRIPTION; Physical Mapping DNA molecules DNA fibers fluorescence in situ hybridization FISH digital image analysis

Citation Formats

Lu, Chun-Mei, Wang, Mei, Greulich-Bode, Karin M, Weier, Jingly F, and Weier, Heinz-Ulli G. Quantitive DNA Fiber Mapping. United States: N. p., 2008. Web.
Lu, Chun-Mei, Wang, Mei, Greulich-Bode, Karin M, Weier, Jingly F, & Weier, Heinz-Ulli G. Quantitive DNA Fiber Mapping. United States.
Lu, Chun-Mei, Wang, Mei, Greulich-Bode, Karin M, Weier, Jingly F, and Weier, Heinz-Ulli G. 2008. "Quantitive DNA Fiber Mapping". United States. https://www.osti.gov/servlets/purl/948493.
@article{osti_948493,
title = {Quantitive DNA Fiber Mapping},
author = {Lu, Chun-Mei and Wang, Mei and Greulich-Bode, Karin M and Weier, Jingly F and Weier, Heinz-Ulli G},
abstractNote = {Several hybridization-based methods used to delineate single copy or repeated DNA sequences in larger genomic intervals take advantage of the increased resolution and sensitivity of free chromatin, i.e., chromatin released from interphase cell nuclei. Quantitative DNA fiber mapping (QDFM) differs from the majority of these methods in that it applies FISH to purified, clonal DNA molecules which have been bound with at least one end to a solid substrate. The DNA molecules are then stretched by the action of a receding meniscus at the water-air interface resulting in DNA molecules stretched homogeneously to about 2.3 kb/{micro}m. When non-isotopically, multicolor-labeled probes are hybridized to these stretched DNA fibers, their respective binding sites are visualized in the fluorescence microscope, their relative distance can be measured and converted into kilobase pairs (kb). The QDFM technique has found useful applications ranging from the detection and delineation of deletions or overlap between linked clones to the construction of high-resolution physical maps to studies of stalled DNA replication and transcription.},
doi = {},
url = {https://www.osti.gov/biblio/948493}, journal = {},
number = ,
volume = ,
place = {United States},
year = {Mon Jan 28 00:00:00 EST 2008},
month = {Mon Jan 28 00:00:00 EST 2008}
}

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