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

Overcoming a barrier for DNA polymerization in triplex-forming sequences.

V N Potaman1, J J Bissler

  • 1Institute of Biosciences and Technology, The Texas A&M University System Health Science Center, 2121 West Holcombe Boulevard, Houston, TX 77030, USA. vpotaman@ibt.tamu.edu

Nucleic Acids Research
|August 24, 1999
PubMed
Summary

DNA polymerases stall on folded DNA templates. A modified PCR method uses an oligonucleotide to prevent template folding, enabling DNA synthesis and analysis of difficult sequences.

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

Effect of everolimus on skin lesions in patients treated for subependymal giant cell astrocytoma and renal angiomyolipoma: final 4-year results from the randomized EXIST-1 and EXIST-2 studies.

Journal of the European Academy of Dermatology and Venereology : JEADV·2018
Same author

Tuberous sclerosis complex: neurological, renal and pulmonary manifestations.

Neuropediatrics·2011
Same author

DNA-binding properties of poly(ADP-ribose) polymerase: a target for anticancer therapy.

Current drug targets·2004
Same author

Length-dependent structure formation in Friedreich ataxia (GAA)n*(TTC)n repeats at neutral pH.

Nucleic acids research·2004
Same author

The structure of intramolecular triplex DNA: atomic force microscopy study.

Journal of molecular biology·2002
Same author

Hematuria in children.

Pediatric clinics of North America·2001

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Folded DNA structures like hairpins and triplexes impede DNA polymerase activity.
  • Mirror-repeated homopurine.homopyrimidine sequences are prone to forming triple-stranded structures, hindering DNA synthesis.

Purpose of the Study:

  • To develop a linear PCR amplification method for structural DNA analysis in challenging mirror-repeated sequences.
  • To overcome DNA synthesis barriers caused by template folding.

Main Methods:

  • Modification of a conventional PCR protocol.
  • Utilizing a synthetic oligonucleotide to hybridize with the DNA template.
  • The oligonucleotide prevents template folding and is displaced by DNA polymerase.

Main Results:

Related Experiment Videos

  • Successfully eliminated the barrier to DNA synthesis on folded DNA templates.
  • Enabled linear PCR amplification for structural analysis of difficult DNA sequences.

Conclusions:

  • The modified PCR approach effectively overcomes DNA polymerase arrest sites caused by DNA folding.
  • This method is valuable for DNA sequencing and analyzing chemical adducts and mutations in sequences prone to forming hairpins and quadruplexes.