PMID: 9171107Jun 15, 1997Paper

A closed tube format for amplification and detection of DNA based on energy transfer

Nucleic Acids Research
I A NazarenkoR J Hohman

Abstract

A new method for the direct detection of PCR-amplified DNA in a closed system is described. The method is based on the incorporation of energy transfer-labeled primers into the amplification product. The PCR primers contain hairpin structures on their 5'ends with donor and acceptor moieties located in close proximity on the hairpin stem. The primers are designed in such a way that a fluorescent signal is generated only when the primers are incorporated into an amplification product. A signal to background ratio of 35:1 was obtained using the hairpin primers labeled with fluorescein as a donor and 4-(4'-dimethylaminophenylazo) benzoic acid (DABCYL) as a quencher. The modified hairpin-primers do not interfere with the activity of DNA polymerase, and both thermostable Pfu and Taq polymerase can be used. This method was applied to the detection of cDNA for prostate specific antigen. The results demonstrate that the fluorescent intensity of the amplified product correlates with the amount of incorporated primers, and as few as 10 molecules of the initial template can be detected. This technology eliminates the risk of carry-over contamination, simplifies the amplification assay and opens up new possibilities for the real-time quanti...Continue Reading

References

Jan 1, 1992·Methods in Enzymology·R M Clegg
Aug 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·P M HollandD H Gelfand
Dec 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·R A CardulloD E Wolf
Jan 1, 1987·Methods in Enzymology·K B Mullis, F A Faloona
Jan 1, 1995·Annual Review of Biophysics and Biomolecular Structure·G Varani
Jun 1, 1993·Trends in Genetics : TIG·U Landegren
Jan 1, 1995·Methods in Enzymology·P R Selvin
May 9, 1995·Proceedings of the National Academy of Sciences of the United States of America·J JuR A Mathies
Aug 11, 1993·Nucleic Acids Research·L G LeeW Bloch

❮ Previous
Next ❯

Citations

Dec 13, 2000·Luminescence : the Journal of Biological and Chemical Luminescence·J NurmiA Ylikoski
Jun 26, 2001·Journal of Biochemical and Molecular Toxicology·N J Walker
Apr 18, 2007·Analytical and Bioanalytical Chemistry·Deming KongHanxi Shen
Nov 4, 2010·Analytical and Bioanalytical Chemistry·Bernard Juskowiak
Dec 2, 2005·Clinical & Experimental Metastasis·Tracey A MartinWen G Jiang
Jan 10, 2002·Infectious Disease Clinics of North America·D WolkR Patel
Jun 17, 1998·Research in Immunology·J P Levraud
Feb 13, 2001·Current Opinion in Biotechnology·B Schweitzer, S Kingsmore
Jan 16, 1999·Current Opinion in Biotechnology·D WhitcombeS Little
Oct 17, 2002·Trends in Parasitology·Andrew S Bell, Lisa C Ranford-Cartwright
Sep 18, 2010·Journal of the American Chemical Society·Michael E ØstergaardPatrick J Hrdlicka
Apr 2, 2002·Organic Letters·Kenneth L SmithSidney M Hecht
Dec 6, 2001·Nature Reviews. Genetics·A C Syvänen
Dec 17, 2005·Chemical Communications : Chem Comm·Rohan T Ranasinghe, Tom Brown
Jun 6, 2007·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Branko KolaricFrans C De Schryver
Feb 2, 2011·Chemical Communications : Chem Comm·Rohan T Ranasinghe, Tom Brown
Aug 29, 2012·The Journal of Biological Chemistry·B Sina Meyer, Jörg Rademann
Dec 24, 2004·Critical Reviews in Food Science and Nutrition·Virginia García-CañasRamón González
Oct 28, 2006·Nucleosides, Nucleotides & Nucleic Acids·C J RankinT P Shields
Aug 14, 1999·Nucleic Acids Research·J G HarrisonS Balasubramanian
Mar 9, 2002·Nucleic Acids Research·Ian M MackayAndreas Nitsche
Apr 25, 2002·Nucleic Acids Research·Irina NazarenkoAyoub Rashtchian

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.