BIOMAT Database Logo BIOMAT Database Logo

Detection of isozymes of deoxyribonucleases I and II on electrophoresed gels with picogram sensitivity using SYBR Green I. 1998

R Iida, and T Yasuda, and E Tsubota, and Y Nakashima, and K Sawazaki, and M Aoyama, and T Matsuki, and K Kishi
Department of Forensic Medicine, Fukui Medical University, Japan.

A highly sensitive method for detecting deoxyribonucleases (DNases) I and II on an electrophoresed gel is described. A dried agarose film sheet containing DNA as a substrate and a buffer reagent was placed in contact with the gel surface after electrophoresis (DAFO method, Yasuda et al., Anal. Biochem. 1989, 183, 84-88). After an appropriate incubation period, the film sheet was peeled off and stained with SYBR-Green I (SG), and then the DNase isozyme bands were detected using a fluorescence image analyzer. We could detect pg levels of the DNases (DNase I, 2 pg; DNase II, 2pg), which represents a 32- to 128-fold increase in sensitivity compared with the original DAFO method using ethidium bromide (EB) as the fluorescent dye. A combination of this new detection method and isoelectric focusing electrophoresis in polyacrylamide gel allowed accurate DNase I typing from 1 microL human serum. This new technique has been named SG-DAFO, after its original dried agarose film overlay method using EB (EB-DAFO).

UI MeSH Term Description Entries
D007525 Isoelectric Focusing Electrophoresis in which a pH gradient is established in a gel medium and proteins migrate until they reach the site (or focus) at which the pH is equal to their isoelectric point. Electrofocusing,Focusing, Isoelectric
D007527 Isoenzymes Structurally related forms of an enzyme. Each isoenzyme has the same mechanism and classification, but differs in its chemical, physical, or immunological characteristics. Alloenzyme,Allozyme,Isoenzyme,Isozyme,Isozymes,Alloenzymes,Allozymes
D008832 Microchemistry The development and use of techniques and equipment to study or perform chemical reactions, with small quantities of materials, frequently less than a milligram or a milliliter.
D009930 Organic Chemicals A broad class of substances containing carbon and its derivatives. Many of these chemicals will frequently contain hydrogen with or without oxygen, nitrogen, sulfur, phosphorus, and other elements. They exist in either carbon chain or carbon ring form. Organic Chemical,Chemical, Organic,Chemicals, Organic
D011804 Quinolines
D003850 Deoxyribonuclease I An enzyme capable of hydrolyzing highly polymerized DNA by splitting phosphodiester linkages, preferentially adjacent to a pyrimidine nucleotide. This catalyzes endonucleolytic cleavage of DNA yielding 5'-phosphodi- and oligonucleotide end-products. The enzyme has a preference for double-stranded DNA. DNase I,Streptodornase,DNA Endonuclease,DNA Nicking Enzyme,DNAase I,Dornavac,Endonuclease I,Nickase,Pancreatic DNase,T4-Endonuclease II,T7-Endonuclease I,Thymonuclease,DNase, Pancreatic,Endonuclease, DNA,T4 Endonuclease II,T7 Endonuclease I
D003959 Diamines Organic chemicals which have two amino groups in an aliphatic chain. Diamine
D004587 Electrophoresis, Agar Gel Electrophoresis in which agar or agarose gel is used as the diffusion medium. Electrophoresis, Agarose Gel,Agar Gel Electrophoresis,Agarose Gel Electrophoresis,Gel Electrophoresis, Agar,Gel Electrophoresis, Agarose
D004591 Electrophoresis, Polyacrylamide Gel Electrophoresis in which a polyacrylamide gel is used as the diffusion medium. Polyacrylamide Gel Electrophoresis,SDS-PAGE,Sodium Dodecyl Sulfate-PAGE,Gel Electrophoresis, Polyacrylamide,SDS PAGE,Sodium Dodecyl Sulfate PAGE,Sodium Dodecyl Sulfate-PAGEs
D004706 Endodeoxyribonucleases A group of enzymes catalyzing the endonucleolytic cleavage of DNA. They include members of EC 3.1.21.-, EC 3.1.22.-, EC 3.1.23.- (DNA RESTRICTION ENZYMES), EC 3.1.24.- (DNA RESTRICTION ENZYMES), and EC 3.1.25.-.

Related Publications

R Iida, and T Yasuda, and E Tsubota, and Y Nakashima, and K Sawazaki, and M Aoyama, and T Matsuki, and K Kishi
Activity measurement for deoxyribonucleases I and II with picogram sensitivity based on DNA/SYBR Green I fluorescence.
January 1998, Analytical biochemistry,
R Iida, and T Yasuda, and E Tsubota, and Y Nakashima, and K Sawazaki, and M Aoyama, and T Matsuki, and K Kishi
Detection of DNA using a visible dye, Nile blue, in electrophoresed gels.
May 2000, Analytical biochemistry,
R Iida, and T Yasuda, and E Tsubota, and Y Nakashima, and K Sawazaki, and M Aoyama, and T Matsuki, and K Kishi
Simple detection of point mutations in DNA oligonucleotides using SYBR Green I.
October 2003, Biotechnology letters,
R Iida, and T Yasuda, and E Tsubota, and Y Nakashima, and K Sawazaki, and M Aoyama, and T Matsuki, and K Kishi
Detection of Plasmodium knowlesi using recombinase polymerase amplification (RPA) combined with SYBR Green I.
August 2020, Acta tropica,
R Iida, and T Yasuda, and E Tsubota, and Y Nakashima, and K Sawazaki, and M Aoyama, and T Matsuki, and K Kishi
Detection of coralyne and heparin by polymerase extension reaction using SYBR Green I.
August 2019, Molecular and cellular probes,
R Iida, and T Yasuda, and E Tsubota, and Y Nakashima, and K Sawazaki, and M Aoyama, and T Matsuki, and K Kishi
[Deoxyribonucleases I and II].
December 1973, Revista espanola de las enfermedades del aparato digestivo,
R Iida, and T Yasuda, and E Tsubota, and Y Nakashima, and K Sawazaki, and M Aoyama, and T Matsuki, and K Kishi
Fluorescent Probe for Ag+ Detection Using SYBR GREEN I and C-C Mismatch.
December 2020, Biosensors,
R Iida, and T Yasuda, and E Tsubota, and Y Nakashima, and K Sawazaki, and M Aoyama, and T Matsuki, and K Kishi
SYBR green I DNA staining increases the detection sensitivity of viruses by polymerase chain reaction.
September 1995, Journal of virological methods,
R Iida, and T Yasuda, and E Tsubota, and Y Nakashima, and K Sawazaki, and M Aoyama, and T Matsuki, and K Kishi
Rapid visual detection of anisakid nematodes using recombinase polymerase amplification and SYBR Green I.
January 2022, Frontiers in microbiology,
R Iida, and T Yasuda, and E Tsubota, and Y Nakashima, and K Sawazaki, and M Aoyama, and T Matsuki, and K Kishi
Quantitation of HTLV-I and II proviral load using real-time quantitative PCR with SYBR Green chemistry.
December 2004, Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology,
Copied contents to your clipboard!