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

Calcium phosphate transfection.

R E Kingston1, C A Chen, H Okayama

  • 1Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA.

Current Protocols in Immunology
|April 25, 2008
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

Decadal stability in coral cover could mask hidden changes on reefs in the East Asian Seas.

Communications biology·2023
Same author

Longitudinal multicentre retrospective cohort study of treatment outcomes in extramammary Paget disease.

The British journal of dermatology·2021
Same author

Multiple introductions and secondary dispersion of Tubastraea spp. in the Southwestern Atlantic.

Scientific reports·2019
Same author

Application of xenogeneic anti-canine distemper virus antibodies in treatment of canine distemper puppies.

The Journal of small animal practice·2016
Same author

Medical Costs of Cardiovascular Diseases in Taiwan.

Value in health : the journal of the International Society for Pharmacoeconomics and Outcomes Research·2016
Same author

Continuous positive airway pressure-associated cutaneous amoebiasis in an immunosuppressed patient.

The British journal of dermatology·2015
Same journal

A Comprehensive Experimental Guide to Studying Cross-Presentation in Dendritic Cells In Vitro.

Current protocols in immunology·2020
Same journal

Immunologic Studies in Humans.

Current protocols in immunology·2020
Same journal

Immunologic Studies in Humans.

Current protocols in immunology·2020
Same journal

Immunologic Studies in Humans.

Current protocols in immunology·2020
Same journal

Monoclonal Antibodies to Human Cell Surface Antigens.

Current protocols in immunology·2020
Same journal

Protocols for Experimental Sjögren's Syndrome.

Current protocols in immunology·2020
See all related articles

This study presents two calcium phosphate-based methods for eukaryotic cell transfection, suitable for transient and stable gene introduction. These protocols optimize plasmid DNA delivery into cell cultures using precipitate formation for enhanced transformation efficiency.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biotechnology

Background:

  • Eukaryotic cell transfection is crucial for genetic studies and biotechnology.
  • Calcium phosphate precipitation is a common method for introducing nucleic acids into cells.
  • Optimizing transfection efficiency and stability is essential for various applications.

Purpose of the Study:

  • To describe two distinct calcium phosphate-based protocols for eukaryotic cell transfection.
  • To provide methods for both transient and stable transfection.
  • To detail procedures for preparing high-quality plasmid DNA and enhancing transfection efficiency.

Main Methods:

  • Basic Protocol: Direct layering of a HEPES-buffered calcium phosphate precipitate onto monolayer cell cultures.

Related Experiment Videos

  • Alternate High-Efficiency Method: Gradual formation of a BES-buffered calcium phosphate precipitate in the medium before application to cells.
  • Support Protocols: Preparation of high-quality plasmid DNA and glycerol or dimethyl sulfoxide (DMSO) shock for increased efficiency.

    • Main Results:

    • Both methods effectively introduce plasmid DNA into eukaryotic cells via precipitate adherence.
    • The alternate BES-buffered method demonstrates high efficiency, particularly for stable transformation with circular plasmid DNA.
    • Transfection efficiency can be further improved using glycerol or DMSO cell shocking.

    Conclusions:

    • Two reliable calcium phosphate-based transfection methods are provided for diverse eukaryotic cell applications.
    • The alternate method offers enhanced efficiency for stable transformation, applicable to various DNA types.
    • High-quality reagents and optimized conditions are key for successful transfection outcomes.