Quantitative DNA fiber mapping

H. U G Weler, Lin Wang, J. C. Mullikin, Y. Zhu, J. F. Cheng, K. M. Greulich, A. Bensimon, Joe Gray

Research output: Contribution to journalArticle

92 Citations (Scopus)

Abstract

The assembly of sequence ready, high-resolution physical maps and construction of minimally overlapping contigs for the human as well as model genomes requires accurate determination of the extent of overlap between adjacent clones as well as their relative orientation. This is presently done by procedures such as clone fingerprinting, Southern blot analysis or clone end sequencing. We present a complementary analytical technique to map directly cloned DNA sequences on to individual stretched DNA molecules. This approach uses the hydrodynamic force of a receding meniscus to prepare straight high molecular weight DNA molecules that provide a linear template of ∼2.3 kb/μm on to which the cloned probes can be mapped by in situ hybridization. This technique has numerous advantages such as a very high density of mapping templates, reproducible stretching of the mapping template providing a linear genomic scale, determination of clone orientation and direct visualization of DNA repeats. The utility and accuracy of quantitative DNA fiber mapping are illustrated through three examples: (i) mapping of lambda DNA restriction fragments along linearized ∼49 kb long lambda phage DNA molecules with ∼1 kb precision; (ii) localization of the overlap between a cosmid and a colinear P1 clone; and (iii) mapping of P1 clones along a ∼490 kb yeast artificial chromosome (YAC) with ∼5 kb precision and estimation of the ∼25 kb gap between them.

Original languageEnglish (US)
Pages (from-to)1903-1910
Number of pages8
JournalHuman Molecular Genetics
Volume4
Issue number10
DOIs
StatePublished - Oct 1995
Externally publishedYes

Fingerprint

Clone
DNA
Clone Cells
Fiber
Fibers
Template
Molecules
Overlap
Yeast Artificial Chromosomes
Bacteriophage lambda
Cosmids
Chromosomes, Human, Pair 5
Bacteriophages
DNA sequences
In Situ Hybridization
Hydrodynamics
Chromosomes
Southern Blotting
Fingerprinting
Yeast

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Public Health, Environmental and Occupational Health
  • Molecular Biology
  • Genetics(clinical)
  • Genetics

Cite this

Weler, H. U. G., Wang, L., Mullikin, J. C., Zhu, Y., Cheng, J. F., Greulich, K. M., ... Gray, J. (1995). Quantitative DNA fiber mapping. Human Molecular Genetics, 4(10), 1903-1910. https://doi.org/10.1093/hmg/4.10.1903

Quantitative DNA fiber mapping. / Weler, H. U G; Wang, Lin; Mullikin, J. C.; Zhu, Y.; Cheng, J. F.; Greulich, K. M.; Bensimon, A.; Gray, Joe.

In: Human Molecular Genetics, Vol. 4, No. 10, 10.1995, p. 1903-1910.

Research output: Contribution to journalArticle

Weler, HUG, Wang, L, Mullikin, JC, Zhu, Y, Cheng, JF, Greulich, KM, Bensimon, A & Gray, J 1995, 'Quantitative DNA fiber mapping', Human Molecular Genetics, vol. 4, no. 10, pp. 1903-1910. https://doi.org/10.1093/hmg/4.10.1903
Weler HUG, Wang L, Mullikin JC, Zhu Y, Cheng JF, Greulich KM et al. Quantitative DNA fiber mapping. Human Molecular Genetics. 1995 Oct;4(10):1903-1910. https://doi.org/10.1093/hmg/4.10.1903
Weler, H. U G ; Wang, Lin ; Mullikin, J. C. ; Zhu, Y. ; Cheng, J. F. ; Greulich, K. M. ; Bensimon, A. ; Gray, Joe. / Quantitative DNA fiber mapping. In: Human Molecular Genetics. 1995 ; Vol. 4, No. 10. pp. 1903-1910.
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