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

Efficient cDNA cloning method using solid phase DNA probe.

H Tsurui1, N Kim, Y Umezawa

  • 1Kanagawa Academy of Science and Technology, Japan.

Nucleic Acids Symposium Series
|January 1, 1992
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

Optimal clinicogenetic criteria for post-operative re-irradiation in recurrent glioblastoma: KROG 21-02.

ESMO open·2026
Same author

Generation of a quasi-Bessel beam with a super-Gaussian axial intensity profile.

Applied optics·2026
Same author

Accuracy of artificial intelligence-assisted growth prediction in skeletal Class I preadolescent patients using serial lateral cephalograms for a 2-year growth interval.

Orthodontics & craniofacial research·2024
Same author

Adrenalectomy for the treatment of hypotension in a cat with phaeochromocytoma associated with caudal vena cava syndrome.

The Journal of small animal practice·2024
Same author

OsCM regulates rice defence system in response to UV light supplemented with drought stress.

Plant biology (Stuttgart, Germany)·2023
Same author

Improved Performance of Organic Thermoelectric Generators Through Interfacial Energetics.

Advanced science (Weinheim, Baden-Wurttemberg, Germany)·2023
Same journal

Nucleotide specificity of DNA cleavage by esperamycin/calichemicin antitumor antibiotics.

Nucleic acids symposium series·2018
Same journal

A model reaction for the repair of nucleic acids. The repair of thymidine bromohydrins.

Nucleic acids symposium series·2018
Same journal

Effects of base damages on DNA replication.

Nucleic acids symposium series·2018
Same journal

Effect of serine residue on the effectiveness of cationic polypeptide-based gene delivery.

Nucleic acids symposium series·2003
Same journal

Smart probe: a novel fluorescence quenching-based oligonucleotide probe carrying a fluorophore and an intercalator.

Nucleic acids symposium series·2003
Same journal

The engineering, structure, and DNA binding properties of a novel His4-type zinc finger peptide.

Nucleic acids symposium series·2003
See all related articles

This study introduces a novel solid-phase DNA probe method for efficiently isolating specific recombinant complementary DNA (cDNA) from single-stranded cDNA libraries. This technique rapidly identifies target clones, like the metapyrocatechase gene, from human lymphoma cell lines.

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetics

Background:

  • Isolating specific recombinant complementary DNA (cDNA) from complex libraries is crucial for gene cloning and functional studies.
  • Traditional methods can be time-consuming and less efficient, particularly with single-stranded cDNA libraries.

Purpose of the Study:

  • To develop and validate a novel, efficient method for isolating target cDNA clones using a solid-phase DNA probe.
  • To demonstrate the method's efficacy in isolating the metapyrocatechase (MPC) gene from a human lymphoma cell line cDNA library.

Main Methods:

  • Conversion of target clone and cDNA library to single-stranded form using helper phage M13KO7.
  • Synthesis of a 25-mer DNA probe from the target cDNA sequence.
  • Immobilization of the DNA probe onto HPLC gel for solid-phase hybridization.

Related Experiment Videos

  • Hybridization, elution, and transformation of competent cells (JM109) using Hanahan's method.

    • Main Results:

    • Successful hybridization and elution of the target clone from the solid-phase DNA probe within hours.
    • Isolation of 50 colonies containing the metapyrocatechase (MPC) gene from a mixture of MPC vector and cDNA library.
    • High efficiency in identifying target colonies from a complex cDNA library.

    Conclusions:

    • The developed solid-phase DNA probe method is a rapid and efficient technique for isolating specific recombinant cDNA.
    • This method offers a significant improvement over conventional techniques for screening cDNA libraries.
    • The approach is effective for isolating genes such as metapyrocatechase from human cell line mRNA.