Nucleic acid labelling

In situ hybridisation Localizes specific genes within cells with nucleic acid-labelled probes Sensitivity and specificity dependent upon tissue and probe quality Yes... 

Fluorescein and its derivatives are one set of fluorescent labels that are commonly used to monitor DNA hybridization, and are often used as labels but do not selectively associate with the structure of double-stranded DNA. The most popular fluorescein derivative for nucleic acid labelling is FAM (car-boxyfluorescein). The structure of FAM is shown in Fig. 3. 

DNA labeled with ULYSIS Alexa Fluor 488, using a Nucleic Acid Labeling Kit (Invitrogen). DNA provided in the kit or oligonucleotides from other suppliers (e.g., Sigma) can be used. 

Nucleic Acids Labeling Strategies Selected Fluorescence Techniques... 

Table 7.4 summarizes the characteristics of radionuclides most often used in nucleic acid labeling. Commercial dNTP preparations usually contain 10 (xCi/jxl and those with higher specific activities thus have a lower concentration. For instance, if the final radioactive concentration in the mixture is 1 jjcCi/p-I, the concentration of that radiolabeled dNTP is 1 piM if the specific activity is 1000 Ci/mmol but 0.2 p,M if the specific activity is 5000 Ci/mmol. 

Characteristics of radioisotopes used in nucleic acid labeling... 

Amersham (1987) Nucleic Acid Labelling. Amersham Corp. Arlington Heights, IL. 51 pp. 

Description of labeHng protocol can include label used (e.g., Cy3, Cy5,33P), amount of nucleic acids labeled, the labeling ratio (efficiency). 

Nucleic acid labelling, detection and quantitation with single fluorescent dye... 

Enzyme-Labeled Fluorescence (ELF) Phosphatase-based signal amplification assay also applicable to nucleic acid labeling (see description under proteins) 43... 

Polyplex uptake is typically assessed by transfecting cells with polyplexes formed with nucleic acids labeled with a fluorescent tag (such as rhodamine, fluorescein, and the alexa fluor dasses of dyes). " In some cases, the polymer or dendrimer can also be labeled with a fluorophore and cellular internalization of both the polymer and the nudeic acid is monitored. The assessment is then typically made by one or more of the following techniques measurement of the cell lysate fluorescence, fluorescence microscopy (wide-field or confocal miaoscopy)... 

Reagents 1 and 2 are available in kit form including the reagents for hybridization and chemiluminescent signal generation (ECL direct nucleic acid labeling and detection system, RPN 3000, RPN 3001, RPN 3005, Amersham International, Amersham, UK). These... 

Nucleic acid labeling reagent charge modified HRP (Amersham). 

Proteins, peptides, and nucleic acid labeling in molecular biology research. The fluorescence modification of nucleic acid molecules can be achieved in a number of ways. For example, fluorescent DNA... 

Figure 1 Molecular structures of some intercalative dyes for nucleic acids labeling.

Ethidium bromide 526 605 0.52 Classic general-purpose nucleic acid label... 

Tintcheva I, Maximova V, DeUgeorgiev T, Zaneva D, Ivanov I. New asymmetric monomethine cyanine dyes for nucleic-acid labelling absorption and fluorescence spectral characteristics, J. Photochem. Photobiol. A Chem. 2000. p. 7 - 11. DOI 10.1016/S1010-6030(99)00207-5... 

Nucleic Acid Labeling and Detection by iEDDA Bioorthogonal Reaction... 

Fig. 1 Cycloaddition reactions employed in nucleic acid labeling with reporter groups (green star). A Cu -mediated azide-alkyne cycloaddition (CuAAC) of a terminal alkyne with an azide. B Strain-promoted azide-alkyne cycloaddition (SPAAC) of an azide with a cyclooctyne derivative. C Staudinger ligation of an azide with a phosphine derivative (not a cycloaddition reaction, see below). D Norbornene cycloaddition of a nitrile oxide as 1,3-dipole and a norbornene as dipolarophile. E Inverse electron-demand Diels- Alder cycloaddition reaction between a strained double bond (norbornene) and a tetrazine derivative. F Photo-cUck reaction of a push-pull-substituted diaiyltetrazole with an activated double bond (maleimide)...

Puromycin Esdierichia colt ribosomes Both protein and nucleic acid labeled 30... 

They are usually synthesised with reactive groups, R, on either one or both of the nitrogen atoms so that they can be chemically linked to nucleic acids or proteins. Used mainly for protein and nucleic acid labelling. Cy3 and Cy5 are commonly combined for 2 colour detection Cy3 dyes fluoresce yellow-green while Cy5 dyes fluoresce red. 

Chemical modification of nucleic acids with functionalized linkers, such as amine-functionalized alkyl chains tethered to nucleic acids, has been employed to modify surfaces with DNA probes [26]. Conductive supports and particularly Au-surfaces were functionaUzed with nucleic acid labeled with an oUgothymine thio-phosphate [(Ts ) ] [27] or with a thiol-functionalized linker tethered to the probe... 

Hoeltke, H. J. Ettl, I. Finken, M. West, S. Kunz, W. Multiple nucleic acid labeling and rainbow detection. Anal. Biochem. 1992,207,24-31. 

Henegariu, O. Bray-Ward, R Ward, D. C. Custom fluorescent-nucleotide synthesis as an alternative method for nucleic acid labeling. Nat. Biotechnol. 2000,18, 345-348. 

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