A well established transfection reagent for mammalian cells, DOTAP™ is a ready to use transfection reagent. It is a liposome formulation of a monocationic lipid (N-[1-(2,3-Dioleoyloxy)]-N,N,N-trimethylammonium propane methylsulphate in sterile water (concentration 1mg/ml).
There are a large number of reports in literature using this reagent (in vitro & in vivo). It combines reliability and efficiency with low costs. 1ml of DOTAP™ is usually sufficient for 50-100 transfections on 35mm dishes (6-well plates). DOTAP™ transfects nucleic acids both in the presence and absence of serum.
If stored at 4°C it is stable for at least 6 months
Structure of DOTAP™
(C43H83NO8S, MW: 774,21)
First-generation transfection reagents
The
unsatisfactory and unreliable results of the calcium phosphate
transfection method led to the development of transfection reagents of
the 1. Generation based on cationic lipids or liposomes.
These
reagents are generally based on monocationic lipids with a quaternary
ammonium ion. They do not feature buffering properties and therefore do
not offer the option of endosome buffering, which explains their lower
efficiency compared to modern reagents (e.g. METAFECTENE®). In addition, serum inhibition is reported for many of these transfection reagents with the exception of DOTAP.
A
further feature of these reagents is a generally conventional
transfection sequence providing for cell seeding and growth prior to the
actual transfection and for change of medium after transfection. As a
rule the proportion of DNA to lipids must be optimized carefully to
ensure satisfactory results.
DOTAP was Biontex
the starting point for the development of new transfection reagents. The
temporal development series includes, in particular: METAFECTENE® < METAFECTENE® PRO < K2® Transfection System < K4® Transfection System. With the development degree the probability of a successful transfection increases (see also General Selection Aid).
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