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.

Robert E Kingston1, Claudia A Chen, John K Rose

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

Current Protocols in Molecular Biology
|February 12, 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

Mutual antagonism between PRC1 condensates and SWI/SNF in chromatin regulation.

Molecular cell·2026
Same author

Multiomic analysis of clonal development reveals new regulators of leukemic cell growth.

Genes & development·2026
Same author

Immunization with virus-like vesicle-based COVID-19 vaccine induces robust systemic and mucosal immunity.

NPJ vaccines·2025
Same author

Mutual Antagonism Between PRC1 Condensates and SWI/SNF in Chromatin Regulation.

bioRxiv : the preprint server for biology·2025
Same author

WSTF nuclear autophagy regulates chronic but not acute inflammation.

Nature·2025
Same author

Modularity of PRC1 composition and chromatin interaction define condensate properties.

Molecular cell·2024
Same journal

Nondenaturing Polyacrylamide Gel Electrophoresis: Preparation and Analysis of DNA.

Current protocols in molecular biology·2021
Same journal

Purification and Concentration of DNA from Aqueous Solutions: Preparation and Analysis of DNA.

Current protocols in molecular biology·2021
Same journal

Expression of Proteins Using Semliki Forest Virus Vectors: Protein Expression.

Current protocols in molecular biology·2021
Same journal

Methylation and Uracil Interference Assays for Analysis of Protein-DNA Interactions: DNA-Protein Interactions.

Current protocols in molecular biology·2021
Same journal

Separation of Double- and Single-Stranded Nucleic Acids Using Hydroxylapatite Chromatography: Preparation and Analysis of DNA.

Current protocols in molecular biology·2021
Same journal

Pulsed-Field Gel Electrophoresis: Preparation and Analysis of DNA.

Current protocols in molecular biology·2021
See all related articles

Two calcium phosphate-based methods efficiently transfect eukaryotic cells for transient and stable expression. These protocols introduce plasmid DNA via precipitate, with glycerol or DMSO enhancing efficiency in some cases.

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 is essential for successful gene delivery.

Purpose of the Study:

  • To present two calcium phosphate-based methods for eukaryotic cell transfection.
  • To evaluate the efficiency of these methods for both transient and stable transfections.
  • To identify factors influencing transfection success.

Main Methods:

  • Two calcium phosphate precipitation techniques were employed for DNA delivery to monolayer cell cultures.
  • Method 1: Direct layering of a HEPES-buffered calcium phosphate precipitate onto cells.

Related Experiment Videos

  • Method 2: Gradual formation of a BES-buffered precipitate in medium, followed by cell exposure.

    • Main Results:

    • Both methods successfully introduced plasmid DNA into eukaryotic cells.
    • Glycerol or DMSO shock improved transfection efficiency for certain cell types.
    • The BES-buffered method demonstrated high efficiency for stable transformation with circular plasmid DNA.
    • Similar results were observed for linear plasmid or genomic DNA, and transient expression with both methods.

    Conclusions:

    • Calcium phosphate-based transfection offers versatile options for transient and stable gene expression in eukaryotic cells.
    • The choice of buffer system and potential cell shocking can be optimized for specific applications.
    • These protocols provide reliable methods for genetic manipulation of cell cultures.