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

A new PCR based method for the generation of nested deletions

J M Whitcomb1, A Rashtchian, S H Hughes

  • 1ABL-Basic Research Program, NCI-Frederick Cancer Research and Development Center, MD 21702-1201.

Nucleic Acids Research
|August 25, 1993
PubMed
Summary

We developed random primed/anchored-PCR (RPA-PCR), a simple method for amplifying and cloning DNA sequences next to known DNA. This technique is useful for DNA sequencing, promoter analysis, and studying gene structures.

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

HIV latency. Specific HIV integration sites are linked to clonal expansion and persistence of infected cells.

Science (New York, N.Y.)·2014
Same author

Mutations in the human immunodeficiency virus type 1 polypurine tract (PPT) reduce the rate of PPT cleavage and plus-strand DNA synthesis.

Journal of virology·2008
Same author

Molecular cloning of PCR products.

Current protocols in molecular biology·2008
Same author

Natural variation of drug susceptibility in wild-type human immunodeficiency virus type 1.

Antimicrobial agents and chemotherapy·2004
Same author

RNase H cleavage of the 5' end of the human immunodeficiency virus type 1 genome.

Journal of virology·2001
Same author

Inhibition of purified recombinant reverse transcriptase from wild-type and zidovudine-resistant clinical isolates of human immunodeficiency virus type 1 by zidovudine, stavudine, and lamivudine triphosphates.

The Journal of infectious diseases·2001

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Cloning DNA fragments adjacent to known sequences is crucial for various molecular biology applications.
  • Existing methods can be complex or lack efficiency in certain scenarios.

Purpose of the Study:

  • To develop a simple and efficient PCR-based protocol for selective amplification and cloning of adjacent DNA segments.
  • To demonstrate the utility of this method for generating DNA deletions for sequencing and other applications.

Main Methods:

  • Development of a novel PCR-based protocol termed random primed/anchored-PCR (RPA-PCR).
  • Application of RPA-PCR to generate a nested set of deletions for DNA sequencing.

Main Results:

Related Experiment Videos

  • RPA-PCR enables selective amplification and efficient cloning of DNA sequences adjacent to known sequences.
  • The method successfully generated a nested set of evenly spaced deletions suitable for DNA sequencing.

    • Conclusions:

    • RPA-PCR is a versatile and simple technique for various molecular biology applications.
    • Potential applications include analysis of eukaryotic promoters, 5' cDNA extension, and cloning of retroviral/transposon insertion sites.