摘要:

AbstractIn July 1997 Professor Harry Rilbe passed away at the age of 84. Harry Rilbe was an outstanding personality skilled in chemistry, optics and mathematics, and he will be remembered for his important contributions to the development of separation methods, notably moving boundary electrophoresis and isoelectric focusing. The latter method is used for analytical and preparative purposes in thousands of laboratories worldwide. Isoelectric focusing is indispensable for characterization of proteins from humans, animals, plants and microorganisms. Isoelectric focusing is thus useful for the understanding of the function of genes and, accordingly, for progress in the life sciences. A biographical retrospect, “A scientific life with chemistry, optics and mathematics”, was published by Harry Rilbe inElectrophoresis1984,5, 1

ISSN:0173-0835    

DOI:10.1002/elps.1150191002

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractDNA electrophoresis is now a fairly mature technology. Nevertheless, as we approach the 21st century, new ideas are frequently suggested that could lead to a revolution for DNA sequencing and mapping. Here, we review some of the novel concepts that have been studied sinceca.1990. Our review focuses on new separation mechanisms, new sieving matrices and recent conceptual advances.

ISSN:0173-0835    

DOI:10.1002/elps.1150191003

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractThe electrophoretic mobilities of DNA molecules in three different molecular weight ladders were measured in polyacrylamide gels containing different acrylamide concentrations (%T) and cross‐linker ratios (%C), cast and run in Trisacetate‐EDTA (TAE) buffer. The apparent pore radius of each gel was estimated from Ferguson plots of the relative mobiliteis of each of the DNA molecules, using the mobility of the monomer fragment in each molecular weight ladder as the reference mobility. The effective size of each of the DNA molecules was estimated from its radius of gyration. The apparent gel pore radii calculated in this manner ranged from 21 nm in gels containing 10.5 %T, 5%C to 200 nm in gels with 4.6%T, 2%C; similar to the values observed for polyacrylamide gels cast and run in Tris‐borate‐EDTA (TBE) buffer (Holmes and Stellwagen,Electrophoresis1991,12, 612–619). Hence, the effective pore size of polyacrylamide gels is essentially independent of whether the gels are cast and run in TAE or T

ISSN:0173-0835    

DOI:10.1002/elps.1150191004

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractBy using fluorescence recovery after photobleaching (FRAP) and electric birefringence, the migration of single‐stranded DNA in polyacrylamide gels and orientation as a response to an electric pulse were investigated. Electrophoretic mobility is in good agreement with the model of biased reptation including fluctuations. The determination of the electrophoretic mobility in solution, μo, allows an estimation of the gel pore diameter seen by the molecule. As previously observed for double‐stranded DNA, the electric birefringence results from two processes: the alignment of the molecule along the electric field and the elongation of the primitive path in the gel, for long single‐stranded DNA (>2000 bases). The combination of results obtained with the two techniques allows us to propose experimental conditions to improve the separation of single‐stranded DNA with pulsed field te

ISSN:0173-0835    

DOI:10.1002/elps.1150191005

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractIn this article, we extend our recently developed lattice model of gel electrophoresis to periodic three‐dimensional gels made of either isolated obstacles or infinitely long fibers. Exact mobilites are calculated using a much improved numerical method that allows us to treat very large systems. A comparison of the exact mobilities and free available volumes indicates that the main assumption of the Ogston‐Morris‐Rodbard‐Chrambach model (OMRCM), which postulates that the mobility (μ) of charged particles is directly related to the fractional gel volume available to them, is not valid. However, a study of the gel concentration and analyte size dependence of the zero‐field mobility indicates that the OMRCM and the Ferguson plots can indeed be used to obtain useful, semi‐quantitative information about the gel properties. A procedure to study more realistic three‐dimensional gel system

ISSN:0173-0835    

DOI:10.1002/elps.1150191006

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractThe electrophoretic mobility of five single tandem repeats (STRs) and four amplified fragment length polymorphisms (AFLPs) in polyacrylamide gels was tested under denaturing and nondenaturing conditions. Relative anomalous mobility in nondenaturing conditions was found in one AT‐rich AFLP (3′ ApoB) and in two AT‐rich STRs (HUMACTBP2 and HUMF13A1). In these cases, highly anomalous electrophoretic behavior was found even when changes were made in the %T value alone. In such cases typing results were affected by the gel composition. The mobility of these systems was nevertheless normal under denaturing conditions. As a consequence of this study, we recommended that the typing of these systems should only be performed under denaturing conditions and we also recommend the further study of the electrophoretic behavior of repetitive DNA polymorphisms, especially AT‐rich systems, before they are used in forensic or genetic appli

ISSN:0173-0835    

DOI:10.1002/elps.1150191007

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractA method is described which allows accurate nonradioactive sizing of most D11S533 alleles, which are characterized by a highly variable number of 6 bp units. In a small population sample of German origin the heterozygosity of the locus was estimated to be 0.933.

ISSN:0173-0835    

DOI:10.1002/elps.1150191008

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractA large‐scale preparative polyacrylamide gel electrophoresis (PAGE) system for the isolation of high‐purity supercoiled plasmid‐DNA is described. This method should prove suitable for the isolation of large DNA molecules, either plasmid or linear DNA, that is required for the production of transgenic animals, for instance. The efficiency of the method is illustrated by the isolation of the gene for the green fluorescent protein, cloned into a mammalian expression vector and used for transfection of eukaryotic

ISSN:0173-0835    

DOI:10.1002/elps.1150191009

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractThe theory of mass transport coupled to reversible protein interactions forms the basis for computer simulation of the isoelectric focusing behavior of several model systems. These include pH‐dependent conformational transition, carrier ampholyte‐induced interactions and protein‐ligand interactions. The computational results compare favorably with experimental observations. In addition, a method is formulated for an isoelectric focusing procedure which enables determination of intrinsic ligand‐binding constants for statistical binding of a charged ligand, binding to heterogeneous sites, and cooperative

ISSN:0173-0835    

DOI:10.1002/elps.1150191010

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractVarious techniques for chemical separation can be described using a generally applicable theory. There are several schools of thought on how a unified separation science should be developed. The theories described include the mass balance equation (i.e.moving boundary), virtual migration distances, and the use of individual capacity factors.

ISSN:0173-0835    

DOI:10.1002/elps.1150191011

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY