scholarly journals A quantitative luminescence assay for nonradioactive nucleic acid probes.

1996 ◽  
Vol 44 (6) ◽  
pp. 657-660 ◽  
Author(s):  
V V Didenko ◽  
P J Hornsby
Keyword(s):  
Alkaline Phosphatase ◽  
Nucleic Acid ◽  
Nucleic Acids ◽  
Light Emission ◽  
Accurate Determination ◽  
Nucleic Acid Probes ◽  
Radioactive Label ◽  
Luminescence Assay ◽  
Antibody Conjugate

In histochemical work with digoxigenin- or biotin-labeled nucleic acid probes, reproducibility of in situ hybridization depends on accurate measurement of the amount of non-radioactive label being used. We describe a rapid and sensitive assay for nonradioactive label incorporated into nucleic acids employing a luminogenic substrate for alkaline phosphatase, CSPD (disodium 3-(4-methoxyspirol¿1,2-dioxetane-3,2'-(5'-chloro)tricyclo [3.3.1.1(3,7)]decan¿-4-yl)phenyl phosphate). An alkaline phosphatase-antibody conjugate was bound to digoxigenin-labeled nucleic acids spotted on nylon membranes. Light emission from the reaction of the bound alkaline phosphatase with CSPD was measured with a luminometer. This method allows an accurate determination of digoxigenin incorporated into nucleic acid probes in the range of 0.5-500 fmol of nonradioactive label.

Study of the interaction of nucleic acid with europium(III) and CTMAB and determination of nucleic acids at nanogram levels by the second-order scattering

2005 ◽  
Vol 536 (1-2) ◽  
pp. 171-178 ◽  
Author(s):  
Fen Ding ◽  
Huichun Zhao ◽  
Shilv Chen ◽  
Jin Ouyang ◽  
Linpei Jin
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Second Order

Address of the President Lord Blackett, O.M., C.H. at the Anniversary Meeting, 30 November 1970

1971 ◽  
Vol 177 (1046) ◽  
pp. 1-14
Keyword(s):  
Natural Products ◽  
Nucleic Acid ◽  
Nucleic Acids ◽  
Biological Systems ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Theoretical Knowledge ◽  
Three Dimensional Structure ◽  
Subsequent Determination

Todd has made highly significant contributions to the chemistry of natural products, in particular in relation to compounds which play important roles in biological systems. His researches on vitamins B 1 , E and B 12 were most elegant and have had far-reaching implications, but none more so than his structural and synthetic studies in the nucleic acid field. Here he developed methods for the synthesis of the nucleosides and for their phosphorylation; his work on the way they are combined made possible the subsequent determination of the three-dimensional structure of the nucleic acids thereby providing the basis for much of the exciting activity in the nucleotide field today. Todd’s achievements arise out of a rare combination of theoretical knowledge and outstanding experimental skill, with the most judicious exploitation of modern techniques. His work and his quality as an investigator have been widely recognized by biologists as well as by organic chemists.

Address of the President Lord Blackett, O.M., C.H. at the Anniversary Meeting, 30 November 1970

1971 ◽  
Vol 321 (1544) ◽  
pp. 1-14
Keyword(s):  
Natural Products ◽  
Nucleic Acid ◽  
Nucleic Acids ◽  
Biological Systems ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Theoretical Knowledge ◽  
Three Dimensional Structure ◽  
Subsequent Determination

The Copley Medal is awarded to Lord Todd, F. R. S. Todd has made highly significant contributions to the chemistry of natural products, in particular in relation to compounds which play important roles in biological systems. His researches on vitamins B 1 , E and B 12 were most elegant and have had far-reaching implications, but none more so than his structural and synthetic studies in the nucleic acid field. Here he developed methods for the synthesis of the nucleosides and for their phosphorylation; his work on the way they are combined made possible the subsequent determination of the three-dimensional structure of the nucleic acids thereby providing the basis for much of the exciting activity in the nucleotide field today. Todd’s achievements arise out of a rare combination of theoretical knowledge and outstanding experimental skill, with the most judicious exploitation of modern techniques. His work and his quality as an investigator have been widely recognized by biologists as well as by organic chemists.

The use of spectroscopic techniques in the study of DNA stability

2000 ◽  
Author(s):  
John Jr SantaLucia
Keyword(s):  
Nucleic Acid ◽  
Diagnostic Tests ◽  
Accurate Determination ◽  
Spectroscopic Techniques ◽  
Dna And Rna ◽  
Repair Efficiency ◽  
Primary Literature ◽  
Folding Thermodynamics ◽  
Data Analysis Methods

Accurate determination of nucleic acid thermodynamics has become increasingly important in understanding biological function as well as applications in biotechnology and pharmaceuticals. Knowledge of the thermodynamics of DNA hybridization and secondary structure formation is necessary for understanding DNA replication fidelity (1), mismatch repair efficiency (2) and the mechanism of DNA triplet repeat diseases (3). In addition, RNA folding thermodynamics are an important aspect of understanding ribozyme catalysis, as well as understanding the regulation of protein expression, mRNA stability and the mechanism of protein synthesis by the ribosome (4). With the genome sequencing era upon us (5), it will increasingly become important to predict the folding and hybridization thermodynamics of DNA and RNA, so that accurate diagnostic tests for genetic and infectious diseases can be developed. Thus, there is a need to develop a database of accurate thermodynamic parameters for different nucleic acid folding motifs (4). This chapter describes practical aspects of the application of UV absorbance temperature profiles to determine the thermodynamics of nucleic acid structural transitions. Protocols and practical advice are presented for issues not normally addressed in the primary literature but that are crucial for the determination of reliable thermodynamics, such as sequence design, sample preparation, choice of buffer, protocols for determining strand concentrations and mixing strands, design of microvolume cuvettes and cell holder, instrumental requirements, data analysis methods, and sources of error. References to the primary literature and reviews are also provided where appropriate.

scholarly journals Nucleic acid metabolism in the ruminant

1969 ◽  
Vol 23 (3) ◽  
pp. 671-682 ◽  
Author(s):  
A. B. Mcallan ◽  
R. H. Smith
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Acid Solution ◽  
Perchloric Acid ◽  
Trichloroacetic Acid ◽  
Acid Metabolism ◽  
Nucleic Acid Metabolism ◽  
Trichloroacetic Acid Solution ◽  
Rna And Dna

1. Procedures, based on those of Schmidt & Thannhauser (1945) and Schneider (1945), for the extraction and estimation of nucleic acids in bovine digesta were examined in detail.2. Final methods which were suitable for routine determination of RNA and DNA were essentially as follows. Digesta samples were extracted in the cold, first with a solution of trichloroacetic acid in ethanol, then with aqueous trichloroacetic acid solution and finally with lipid solvents. The dried residue was hydrolysed with alkali, purified by passage through a Dowex resin, and the RNA, in the form of mononucleotides, determined by U.V. absorption. DNA was determined separately in hot perchloric acid extracts of the original dried residue by colorimetric estimation of the deoxyribose content.

scholarly journals An impedimetric determination of alkaline phosphatase activity based on the oxidation reaction mediated by Cu2+ bound to poly-thymine DNA

RSC Advances ◽  
2018 ◽  
Vol 8 (20) ◽  
pp. 11241-11246 ◽  
Author(s):  
Joon Young Lee ◽  
Jun Ki Ahn ◽  
Ki Soo Park ◽  
Hyun Gyu Park
Keyword(s):  
Alkaline Phosphatase ◽  
Ascorbic Acid ◽  
Phosphatase Activity ◽  
Modified Electrode ◽  
Alkaline Phosphatase Activity ◽  
Oxidation Reaction ◽  
Accurate Determination ◽  
Alp Activity ◽  
Mediated Oxidation

A novel impedimetric assay for the accurate determination of alkaline phosphatase (ALP) activity is developed based on the Cu2+-mediated oxidation of ascorbic acid on a poly-thymine DNA-modified electrode.

A sensitive dual colorimetric and fluorescence system for assaying the activity of alkaline phosphatase that relies on pyrophosphate inhibition of the peroxidase activity of copper ions

The Analyst ◽  
2014 ◽  
Vol 139 (18) ◽  
pp. 4691-4695 ◽  
Author(s):  
Ki Soo Park ◽  
Chang Yeol Lee ◽  
Hyun Gyu Park
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Peroxidase Activity ◽  
Copper Ions ◽  
Accurate Determination ◽  
Highly Sensitive ◽  
Optical Assay

A novel and highly sensitive optical assay for the accurate determination of alkaline phosphatase activity is developed by utilizing the peroxidase activity of Cu2+ ions.

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