Alignment and sensitive detection of DNA by a moving interface.

In a process called "molecular combining," DNA molecules attached at one end to a solid surface were extended and aligned by a receding air-water interface and left to dry on the surface. Molecular combing was observed to extend the length of the bacteriophage lambda DNA molecule to 21.5 +/- 0.5 micrometers (unextended length, 16.2 micrometers). With the combing process, it was possible to (i) extend a chromosomal Escherichia coli DNA fragment (10(6) base pairs) and (ii) detect a minute quantity of DNA (10(3) molecules). These results open the way for a faster physical mapping of the genome and for the detection of small quantities of target DNA from a population of molecules.

[1]  J. Callis,et al.  Observation of individual DNA molecules undergoing gel electrophoresis. , 1989, Science.

[2]  K. Mullis,et al.  Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. , 1985, Science.

[3]  David C. Schwartz,et al.  Conformational dynamics of individual DNA molecules during gel electrophoresis , 1989, Nature.

[4]  J. Guesdon Immunoenzymatic techniques applied to the specific detection of nucleic acids. A review. , 1992, Journal of immunological methods.

[5]  T. Houseal,et al.  Real-time imaging of single DNA molecules with fluorescence microscopy. , 1989, Biophysical journal.

[6]  R. H. Austin,et al.  DNA electrophoresis in microlithographic arrays , 1992, Nature.

[7]  E. Hornes,et al.  Assessment of methods for covalent binding of nucleic acids to magnetic beads, Dynabeads, and the characteristics of the bound nucleic acids in hybridization reactions. , 1988, Nucleic acids research.

[8]  D. Schwartz,et al.  Ordered restriction maps of Saccharomyces cerevisiae chromosomes constructed by optical mapping. , 1993, Science.

[9]  Robert R. Birge,et al.  Applications of fluorescence in the biomedical sciences , 1986 .

[10]  S. Smith,et al.  Direct mechanical measurements of the elasticity of single DNA molecules by using magnetic beads. , 1992, Science.

[11]  D E Smith,et al.  Direct observation of tube-like motion of a single polymer chain. , 1994, Science.

[12]  J. Guesdon,et al.  Miniaturization of beta-galactosidase immunoassays using chromogenic and fluorogenic substrates. , 1982, Journal of immunological methods.

[13]  J. Langdale,et al.  A rapid method of gene detection using DNA bound to Sephacryl. , 1985, Gene.

[14]  S. Quake,et al.  Relaxation of a single DNA molecule observed by optical microscopy. , 1994, Science.

[15]  F. Rondelez,et al.  Evidence of a transition temperature for the optimum deposition of grafted monolayer coatings , 1992, Nature.

[16]  R E Glass,et al.  Visualization of single molecules of RNA polymerase sliding along DNA. , 1993, Science.

[17]  G Hermanson,et al.  High-resolution mapping of human chromosome 11 by in situ hybridization with cosmid clones. , 1990, Science.

[18]  S. Chu,et al.  Laser Manipulation of Atoms and Particles , 1991, Science.

[19]  E. Cox,et al.  DNA stretching on functionalized gold surfaces. , 1994, Nucleic acids research.

[20]  B. Windle,et al.  High resolution visual mapping of stretched DNA by fluorescent hybridization , 1993, Nature Genetics.