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Fluorescence in situ hybridization method for measuring transfection efficiency.

Publication ,  Journal Article
Cheng, L; Bucana, CD; Wei, Q
Published in: Biotechniques
September 1996

We describe here the use of fluorescence in situ hybridization (FISH) to measure the transfection efficiency of the transient expression vector pCMVcat in lymphoblasts and fibroblasts. By using a pCMVcat probe, we can visualize the location of the plasmid after transfection and thus determine transfection efficiency. In this report, we show that, for transfection of pCMVcat by the diethylaminoethyl-dextran method, the transfection efficiency was about 15 and 70 times greater in fibroblasts and lymphoblasts, respectively, when measured by the FISH method as compared to the efficiency measured by cotransfection with pCMV beta gal. Based on these results, we conclude that the FISH method is a highly sensitive, specific and direct measure of transfection efficiency of a transient expression vector and that it may be useful for evaluating laboratory assays in which the quantitative aspects of transfection and the effect of plasmid DNA damage on transfection efficiency are important.

Duke Scholars

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Published In

Biotechniques

DOI

ISSN

0736-6205

Publication Date

September 1996

Volume

21

Issue

3

Start / End Page

486 / 491

Location

England

Related Subject Headings

  • Xeroderma Pigmentosum
  • Transfection
  • Sensitivity and Specificity
  • Recombinant Fusion Proteins
  • Lymphocytes
  • In Situ Hybridization, Fluorescence
  • Humans
  • Genetic Vectors
  • Genes, Reporter
  • Fibroblasts
 

Citation

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Cheng, L., Bucana, C. D., & Wei, Q. (1996). Fluorescence in situ hybridization method for measuring transfection efficiency. Biotechniques, 21(3), 486–491. https://doi.org/10.2144/96213rr01
Cheng, L., C. D. Bucana, and Q. Wei. “Fluorescence in situ hybridization method for measuring transfection efficiency.Biotechniques 21, no. 3 (September 1996): 486–91. https://doi.org/10.2144/96213rr01.
Cheng L, Bucana CD, Wei Q. Fluorescence in situ hybridization method for measuring transfection efficiency. Biotechniques. 1996 Sep;21(3):486–91.
Cheng, L., et al. “Fluorescence in situ hybridization method for measuring transfection efficiency.Biotechniques, vol. 21, no. 3, Sept. 1996, pp. 486–91. Pubmed, doi:10.2144/96213rr01.
Cheng L, Bucana CD, Wei Q. Fluorescence in situ hybridization method for measuring transfection efficiency. Biotechniques. 1996 Sep;21(3):486–491.

Published In

Biotechniques

DOI

ISSN

0736-6205

Publication Date

September 1996

Volume

21

Issue

3

Start / End Page

486 / 491

Location

England

Related Subject Headings

  • Xeroderma Pigmentosum
  • Transfection
  • Sensitivity and Specificity
  • Recombinant Fusion Proteins
  • Lymphocytes
  • In Situ Hybridization, Fluorescence
  • Humans
  • Genetic Vectors
  • Genes, Reporter
  • Fibroblasts