[10] Enzymes for modifying and labeling DNA and RNA

1987 ◽  
pp. 94-110 ◽  
Author(s):  
Fabio Cobianchi ◽  
Samuel H. Wilson
Keyword(s):  
Dna And Rna

High-resolution nucleic acid sequence mapping via in situ hybridization at the Electron Microscope level

1995 ◽  
Vol 53 ◽  
pp. 752-753
Author(s):  
B.A. Hamkalo ◽  
S. Narayanswami ◽  
A.P. Kausch
Keyword(s):  
Nucleic Acid ◽  
Interphase Nucleus ◽  
Genome Mapping ◽  
Precise Location ◽  
Electron Microscope Level ◽  
Rna Sequences ◽  
Fundamental Interest ◽  
Whole Cells ◽  
Dna And Rna

The availability of nonradioactive methods to label nucleic acids an the resultant rapid and greater sensitivity of detection has catapulted the technique of in situ hybridization to become the method of choice to locate of specific DNA and RNA sequences on chromosomes and in whole cells in cytological preparations in many areas of biology. It is being applied to problems of fundamental interest to basic cell and molecular biologists such as the organization of the interphase nucleus in the context of putative functional domains; it is making major contributions to genome mapping efforts; and it is being applied to the analysis of clinical specimens. Although fluorescence detection of nucleic acid hybrids is routinely used, certain questions require greater resolution. For example, very closely linked sequences may not be separable using fluorescence; the precise location of sequences with respect to chromosome structures may be below the resolution of light microscopy(LM); and the relative positions of sequences on very small chromosomes may not be feasible.

Scanning tunneling microscopy (STM) of DNA and RNA

1989 ◽  
Vol 47 ◽  
pp. 34-35
Author(s):  
Patricia G. Arscott ◽  
Gil Lee ◽  
Victor A. Bloomfield ◽  
D. Fennell Evans
Keyword(s):  
Molecular Structures ◽  
Inorganic Materials ◽  
Resolving Power ◽  
Scanning Tunneling ◽  
Tunneling Microscopy ◽  
Form And Function ◽  
Dna And Rna ◽  
Calf Thymus ◽  
And Function ◽  
The Relationship

STM is one of the most promising techniques available for visualizing the fine details of biomolecular structure. It has been used to map the surface topography of inorganic materials in atomic dimensions, and thus has the resolving power not only to determine the conformation of small molecules but to distinguish site-specific features within a molecule. That level of detail is of critical importance in understanding the relationship between form and function in biological systems. The size, shape, and accessibility of molecular structures can be determined much more accurately by STM than by electron microscopy since no staining, shadowing or labeling with heavy metals is required, and there is no exposure to damaging radiation by electrons. Crystallography and most other physical techniques do not give information about individual molecules.We have obtained striking images of DNA and RNA, using calf thymus DNA and two synthetic polynucleotides, poly(dG-me5dC)·poly(dG-me5dC) and poly(rA)·poly(rU).

Direct Detection of Enteropathogenic Bacteria in Estuarine Water Using Nucleic Acid Probes

1991 ◽  
Vol 24 (2) ◽  
pp. 261-266 ◽  
Author(s):  
Ivor T. Knight ◽  
Jocelyne DiRuggiero ◽  
Rita R. Colwell
Keyword(s):  
Water Samples ◽  
Pathogenic Bacteria ◽  
Direct Detection ◽  
Specific Gene ◽  
Estuarine Water ◽  
Indicator Organisms ◽  
Salmonella Spp ◽  
Dna And Rna ◽  
Enteropathogenic Bacteria ◽  
Highly Sensitive Detection

Direct detection and enumeration of pathogenic bacteria, rather than indicator organisms, in aquatic environments is desirable but hindered by the difficulties of culturing and identifying specific pathogens from these environments. We have developed a method for concentrating bacteria from water samples and extracting their DNA and RNA for use as targets for pathogen-specific gene probes. The method has been used to detect and enumerate Salmonella spp. in estuarine water samples. The probe binds Salmonella DNA quantitatively, making it possible to estimate relative amounts of target in each sample. Salmonella spp. were detected in samples which yielded no Salmonella spp. using culturing. Since the probe method does not require culturing the target organism, both culturable and non-culturable forms are detected. We have also used polymerase chain reaction to amplify a region of the enterotoxin gene in enterotoxigenic Escherichiacoli and Vibriocholerae (ltx and ctx, respectively). The amplified products are then identified with ctx and ltx probes, making specific, highly sensitive detection possible.

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