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

Minimizing DNA recombination during long RT-PCR

G Fang1, G Zhu, H Burger

  • 1Wadsworth Center, New York State Department of Health, Albany 12201, USA.

Journal of Virological Methods
|January 29, 1999
PubMed
Summary
This summary is machine-generated.

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

[Role of intraoperative neurophysiological monitoring in high-risk endovascular aortic repair for spinal cord ischemia].

Zhonghua wai ke za zhi [Chinese journal of surgery]·2026
Same author

[Strategies for aortic arch branch reconstruction during thoracic endovascular aortic repair].

Zhonghua wai ke za zhi [Chinese journal of surgery]·2026
Same author

[Application and value of intravascular ultrasound for excimer laser ablation combined with drug-coated balloon in the treatment of lower limb arteriosclerotic obliterans].

Zhonghua wai ke za zhi [Chinese journal of surgery]·2023
Same author

[Application value of excimer laser ablation combined with drug-coated balloon in non-stent atherosclerotic lesions of lower extremity arteries].

Zhonghua wai ke za zhi [Chinese journal of surgery]·2022
Same author

Melanoma in women of childbearing age and in pregnancy in California, 1994-2015: a population-based cohort study.

Journal of the European Academy of Dermatology and Venereology : JEADV·2022
Same author

Scintillation light detection in the 6-m drift-length ProtoDUNE Dual Phase liquid argon TPC.

The European physical journal. C, Particles and fields·2022

This study minimizes DNA recombination during long reverse transcription polymerase chain reaction (RT-PCR) for HIV-1 research. Optimizing reaction conditions, particularly completing DNA synthesis and adding proofreading, significantly reduces recombination frequency.

Area of Science:

  • Molecular Biology
  • Virology
  • Biotechnology

Background:

  • Advancements enable reverse transcription and amplification of long cDNA, including the HIV-1 genome.
  • Optimizing long RT-PCR requires understanding and minimizing DNA recombination frequency.

Purpose of the Study:

  • To investigate methods for minimizing DNA recombination during long RT-PCR of HIV-1.
  • To identify key factors influencing recombination in combined RT-PCR procedures.

Main Methods:

  • Experimentation with RT conditions, PCR conditions, and template molar ratios.
  • Utilized two distinct HIV-1 strains as templates.
  • Employed strain-specific probes for recombination detection.

Main Results:

Related Experiment Videos

  • Completing DNA strand synthesis during RT is crucial for reducing recombination.
  • Incorporating a proofreading function into the PCR step effectively minimizes recombination.
  • Adjusting reaction parameters allows for control over recombination frequency.

Conclusions:

  • Strategies focusing on complete DNA synthesis and PCR proofreading are most effective in reducing recombination.
  • Recombination frequency in long RT-PCR can be significantly controlled and reduced by optimizing reaction conditions.