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

Bacteriophage lambda cloning system for the construction of directional cDNA libraries.

P S Meissner, W P Sisk, M L Berman

    Proceedings of the National Academy of Sciences of the United States of America
    |June 1, 1987
    PubMed
    Summary

    A new bacteriophage lambda cloning vector, lambda ORF8, enables efficient directional cloning of cDNA libraries. This method simplifies cDNA library construction, reducing RNA and effort requirements for genetic research.

    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

    Malignant eccrine spiradenoma of the vulva: a case report and review of the literature.

    International journal of gynecological cancer : official journal of the International Gynecological Cancer Society·2006
    Same author

    Surgical management of epithelial ovarian cancer: a review of the literature.

    Minerva ginecologica·2005
    Same author

    HPV infection and number of lifetime sexual partners are strong predictors for 'natural' regression of CIN 2 and 3.

    British journal of cancer·2003
    Same author

    A randomized, double blind, Phase III trial using oral beta-carotene supplementation for women with high-grade cervical intraepithelial neoplasia.

    Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology·2001
    Same author

    Vaginal hysterectomy as primary treatment of endometrial cancer in medically compromised women.

    Obstetrics and gynecology·2001
    Same author

    Primary invasive carcinoma of the vagina: treatment with interstitial brachytherapy.

    Cancer·2001

    Area of Science:

    • Molecular Biology
    • Genetics
    • Biotechnology

    Background:

    • Traditional cDNA library construction can be resource-intensive.
    • Directional cloning is crucial for accurate gene expression studies.

    Purpose of the Study:

    • To develop a novel bacteriophage lambda cloning vector, lambda ORF8, for efficient directional cDNA library construction.
    • To establish a streamlined method for preparing cDNAs for directional cloning.

    Main Methods:

    • Modification of the bacteriophage lambda genome to create lambda ORF8 with a unique multiple cloning site.
    • Utilizing oligo(dT) priming and a bifunctional linker for directional cDNA synthesis.
    • Employing HindIII and BamHI endonuclease digestion for directional insertion into lambda ORF8.

    Related Experiment Videos

    Main Results:

    • Construction of a lambda ORF8-based cDNA library from phytohemagglutinin-stimulated human peripheral blood lymphocytes.
    • The primary library contained 2 X 10(8) plaque-forming phage, with over 80% containing inserts.
    • Verification of library representativeness through the identification of gamma-interferon-related clones.

    Conclusions:

    • The lambda ORF8 system provides a simple, efficient method for creating large, directionally cloned cDNA libraries.
    • This approach significantly reduces the amount of RNA and effort needed for library preparation.
    • The developed vector and method are valuable tools for molecular biology and genetic research.