Separation of random fragments of DNA according to properties of their sequences.
TLDR
It is suggested that each DNA molecule undergoes partial melting as it encounters a concentration of denaturants sufficient to melt its least stable sequence, while other sequences remain double stranded; in the partially melted configuration, DNA can continue migration only slowly.Abstract:
The separation of DNA fragments by electrophoresis at high temperature in a denaturing gradient is independent of the length of the fragments. We have suggested that the basis of fragment separation is that each DNA molecule undergoes partial melting as it encounters a concentration of denaturants sufficient to melt its least stable sequence, while other sequences remain double stranded; in the partially melted configuration, DNA can continue migration only slowly. This model is consistent with the observation that fragments of lambda phage DNA cleaved by different restriction endonucleases reach the same final depth in the gel if they contain the same least-stable sequence. A unique set of bands is produced from the electrophoresis of randomly fragmented DNA; this would be expected if there were a limited number of melting centers occupying discrete genetic loci. An intact DNA molecule penetrates about as deeply into the gel as the uppermost band after fragmentation; this would be expected only if the least-stable sequence controls the final depth of the whole molecule.read more
Citations
More filters
Journal ArticleDOI
DNA fragments differing by single base-pair substitutions are separated in denaturing gradient gels: correspondence with melting theory.
S G Fischer,Leonard S. Lerman +1 more
TL;DR: DNA fragments 536 base pairs long differing by single base-pair substitutions were clearly separated in denaturing gradient gel electrophoresis, andcision of the lowest melting domain allows recognition of mutants in the next ranking domain.
Journal ArticleDOI
How sensitive is PCR‐SSCP?
Kenshi Hayashi,David W. Yandell +1 more
TL;DR: The percentage of mutations that can be detected by this technique under various controlled conditions is estimated, and some critical elements affecting sensitivity are described.
Journal ArticleDOI
Why does the electrophoretic mobility of DNA in gels vary with the length of the molecule
Journal ArticleDOI
No Structural Mutation in the Dopamine D2 Receptor Gene in Alcoholism or Schizophrenia: Analysis Using Denaturing Gradient Gel Electrophoresis
Pablo V. Gejman,Anca Ram,Joel Gelernter,Eitan Friedman,Qiuhe Cao,David Pickar,Kenneth Blum,Ernest P. Noble,Henry R. Kranzler,Stephanie O'Malley,Dean H. Hamer,Flanagan Whitsitt,Peter A. Rao,Lynn E. DeLisi,Matti Virkkunen,Markku Linnoila,David Goldman,Elliot S. Gershon +17 more
TL;DR: No structural coding abnormalities in the DRD2 gene are present in alcoholism or schizophrenia, and three infrequent DNA variants that predict altered amino acid sequence of the receptor are found.
Journal ArticleDOI
MDM-2 Oncoprotein Overexpression, p53 Gene Mutation, and VEGF Up-Regulation in Angiosarcomas
Christian Zietz,Matthias Rössle,Christian J. Haas,Andrea Sendelhofert,Astrid Hirschmann,Michael Stürzl,Udo Löhrs +6 more
TL;DR: The data suggest that the p53/ MDM-2 pathway is impaired in about two-thirds of the angiosarcomas and may be a key event in the pathogenesis of human angiosrcomas, and the increased VEGF expression observed supports this hypothesis.
References
More filters
Journal ArticleDOI
Length-independent separation of DNA restriction fragments in two-dimensional gel electrophoresis
TL;DR: The resolving power of the system is demonstrated by the clear resolution of over 250 fragments of the Eco RI digest of E. coli DNA, which provides a two-dimensional separation of fragments when combined with length-dependent electrophoresis in agarose in the perpendicular direction.
Journal ArticleDOI
Action of Nicking-Closing Enzyme on Supercoiled and Nonsupercoiled Closed Circular DNA: Formation of a Boltzmann Distribution of Topological Isomers
TL;DR: It was demonstrated that "nonsupercoiled" closed circular duplex molecules serve as substrates for the nicking-closing enzyme, and that a distribution of topological isomers is generated.
Journal ArticleDOI
A denaturation map of the λ phage DNA molecule determined by electron microscopy
TL;DR: The helix—random coil transition of DNA from λ bacteriophage has been studied by electron microscopy and it is found that the sites occur with high frequency at three zones on a DNA denaturation map.
Journal ArticleDOI
Renaturation Kinetics and thermal stability of DNA in aqueous solutions of formamide and urea
TL;DR: The results of a systematic study of the effects of formamide and urea on the thermal stability and renaturation kinetics of DNA found the dependence of Tm on percent formamide was found to be slightly greater.
Journal ArticleDOI
Denaturation maps of the left and right sides of the lambda DNA molecule determined by electron microscopy
TL;DR: Partially Denatured λ DNA has been examined in the electron microscope and the position of a large number of denatured sites located and expressed in the form of denaturation maps are expressed.
Related Papers (5)
DNA fragments differing by single base-pair substitutions are separated in denaturing gradient gels: correspondence with melting theory.
S G Fischer,Leonard S. Lerman +1 more