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Nucleic Acids Research
Article . 1990 . Peer-reviewed
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Mini-transfections on 96-well microtiter plates

Authors: G, Butticè; M, Kurkinen;

Mini-transfections on 96-well microtiter plates

Abstract

Transient transfection is one of the most commonly used techniques to study gene regulation. Different reporter genes have been used to quantitate the promoter activity. CAT (chloramphenicol acetyltransferase) is the reporter gene most often used in these experiments (1). For increased sensitivity, growth hormone and luciferase have also been used as reporter genes (2, 3). For a rapid analysis of gene expression, we describe here a fast and convenient transfection protocol for growth hormone reporter gene. Because growth hormone is secreted, we have scaled down the number of transfected cells so that cell cultures on 96-well microtiter plates are adequate for routine work. Using this transfection protocol, amounts of DNA, chemicals and nutrients are similarly reduced. The protocol is convenient for multi-variable studies and should facilitate screening for factors and treatments that can result in synergistic stimulation or repression of gene activity. As shown in Fig. 1, results from the mini-transfections are compatible with those obtained by large scale CAT assay or RNase protection technique.

Keywords

Chloramphenicol O-Acetyltransferase, Gene Expression, Metalloendopeptidases, Matrix Metalloproteinase 3, Promoter Regions, Genetic, Transfection

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
5
Average
Average
Average
gold