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High Efficiency Gene Transfer by Multiple Transfection Protocol

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Abstract

A high efficiency transfection protocol employing a common polycationic lipid is described. Using LipofectAMINE, a widely used transfection reagent, we transfected 293T cells with a plasmid harboring the β-galactosidase (β-gal) gene. The transfection efficiency was determined by direct staining for X-gal. The conventional transfection protocol achieved an efficiency of <40% while our protocol, which employs the repetition of transfection a few times, achieved an efficiency of approximately 80%. Thus, a dramatic increase in transfection efficiency can be obtained by simply repeating transfection with the use of a common polycationic lipid. This method will be useful in many molecular biological experiments.

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Authors and Affiliations

  1. Cardiovascular & Pulmonary Research Institute, Allegheny, University of the Health Sciences, Pittsburgh, PA, 15212, USA

    Manabu Yamamoto, Satoshi Okumura, Carsten Schwencke, Junichi Sadoshima & Yoshihiro Ishikawa

Authors
  1. Manabu Yamamoto
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  2. Satoshi Okumura
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  3. Carsten Schwencke
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  4. Junichi Sadoshima
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  5. Yoshihiro Ishikawa
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Yamamoto, M., Okumura, S., Schwencke, C. et al. High Efficiency Gene Transfer by Multiple Transfection Protocol. Histochem J 31, 241–243 (1999). https://doi.org/10.1023/A:1003598614323

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  • DOI: https://doi.org/10.1023/A:1003598614323

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