Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Search research articles

Related Experiment Videos

An adaptor strategy to subclone entire cDNA libraries as single insert recombinants.

¹Medizinische Hochschule Hannover, Institut für Medizinische, Mikrobiologie, FRG.

|October 1, 1989

Summary

This study introduces a novel adaptor strategy to prevent multiple DNA inserts during cDNA library subcloning. This method ensures efficient single-insert recombinants, improving molecular cloning techniques.

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

Revisiting susceptibility testing in MDR-TB by a standardized quantitative phenotypic assessment in a European multicentre study.

The Journal of antimicrobial chemotherapy·2015

Same author

Complement and the regulation of humoral immune responses.

Immunology today·2014

Same author

Transmission of multidrug-resistant tuberculosis in a low-incidence setting, Switzerland, 2006 to 2012.

Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin·2014

Same author

Evaluation of the AID TB resistance line probe assay for rapid detection of genetic alterations associated with drug resistance in Mycobacterium tuberculosis strains.

Journal of clinical microbiology·2014

Same author

The critical influence of the intermediate category on interpretation errors in revised EUCAST and CLSI antimicrobial susceptibility testing guidelines.

Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases·2012

Same author

Mycobacterium tuberculosis population structure determines the outcome of genetics-based second-line drug resistance testing.

Antimicrobial agents and chemotherapy·2012

Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

Subcloning entire cDNA libraries often yields multiple inserts per recombinant.
This is due to self-ligatable DNA fragments released from bulk preparations.
Existing methods lack efficiency in generating single-insert recombinants.

Purpose of the Study:

To develop an efficient method for subcloning entire cDNA libraries with single inserts.
To overcome the limitation of multiple inserts in current subcloning protocols.
To improve the accuracy and efficiency of molecular cloning.

Main Methods:

An adaptor strategy was employed to modify DNA inserts.
Non-compatible ends were conferred to primarily self-ligatable inserts.

Related Experiment Videos

This facilitated efficient library subcloning.

Main Results:

The adaptor strategy successfully prevented multiple inserts in recombinants.
Efficient subcloning of entire cDNA libraries as single-insert recombinants was achieved.
The method enhances the yield of desired single-insert clones.

Conclusions:

The presented adaptor strategy is effective for single-insert cDNA library subcloning.
This method offers a significant improvement over traditional techniques.
It provides a valuable tool for molecular biologists and genetic researchers.