Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Amplification of target-specific, ligation-dependent circular probe

D Y Zhang1, M Brandwein, T C Hsuih

  • 1The Lillian, Henry M. Stratton-Hans Popper Department of Pathology, Department of Otolaryngology, Mount Sinai School of Medicine, New York, NY 10029, USA. david_zhang@smtplink.mssm.edu

Gene
|June 18, 1998
PubMed
Summary

A novel circularizable probe (C-probe) enables super-exponential gene amplification, significantly increasing sensitivity and offering decontamination of carryover amplicons for DNA and RNA detection.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

[Clinical characteristics and prognostic evaluation of patients with hematological disease and sepsis in the Hematological intensive care unit].

Zhonghua xue ye xue za zhi = Zhonghua xueyexue zazhi·2025
Same author

[Immune thrombotic thrombocytopenic purpura treated with Obinutuzumab: a case report and literature review].

Zhonghua xue ye xue za zhi = Zhonghua xueyexue zazhi·2025
Same author

[Analysis of HCV genotype distribution characteristics in the Hotan region].

Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology·2025
Same author

[Study of the occurrence of metabolic dysfunction-associated fatty liver disease in obstructive sleep apnea hypopnea syndrome and its risk factors].

Zhonghua nei ke za zhi·2025
Same author

Evaluation of the effect of botulinum toxin A on the physical and mental health of patients with hemifacial spasm.

Neurologia·2024
Same author

[Application of metabolomics in clinical research on pediatric congenital heart disease].

Zhonghua er ke za zhi = Chinese journal of pediatrics·2024

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetics

Background:

  • Conventional polymerase chain reaction (PCR) methods have limitations in amplification efficiency and sensitivity.
  • Detecting specific nucleic acid sequences, especially at low concentrations, remains a challenge in molecular diagnostics.

Purpose of the Study:

  • To introduce a novel circularizable oligodeoxyribonucleotide probe (C-probe) for enhanced gene amplification.
  • To demonstrate the increased sensitivity and decontamination capabilities of the C-probe method compared to traditional PCR.

Main Methods:

  • Developed a C-probe with target-complementary regions and a primer binding site, enabling ligation to form a circular structure around the target DNA.
  • Utilized rolling circle-like replication and ramification extension amplification for multimeric single-stranded DNA generation.

Related Experiment Videos

  • Employed thermocycling for denaturation and subsequent rounds of primer extension and DNA complex ramification.

    • Main Results:

    • Achieved super-exponential amplification kinetics, surpassing conventional PCR.
    • Demonstrated 1000-fold increased sensitivity in detecting Epstein-Barr virus early RNA compared to linear hemiprobes.
    • Showcased the C-probe's resistance to exonuclease digestion, effectively eliminating carryover amplicon contamination.

    Conclusions:

    • The C-probe method offers significantly improved amplification power and sensitivity for DNA and RNA detection.
    • The ligated C-probe's resistance to exonuclease provides a robust mechanism for decontamination of PCR reactions.
    • The developed technique has broad applicability in molecular diagnostics and research, including a novel isothermal amplification method.