quest for lipofectin protocol

carlos h. pedemonte CPedemonte at UH.EDU
Sat Nov 5 14:40:13 EST 1994



>I'm looking for an alternative to using Life Technologies' (Gibco-BRL)
>lipofectin.  At $130/ml, I feel like I'm getting royally ripped-off,
>especially when I consider the rate at which I use it.  I hear it's not
>difficult to make and the cost of the materials turns out to be about
>$1/ml.  If anybody has a protocol on how to make a lipofectin-like reagent
>could you please send it to me.  Thanks!!!

>               Denis Guttridge
>E-mail: dcguttri at uci.edu
>address: UCI department of microbiology
>         Med Sci I B268
>         Irvine, CA 92717
>        (714)856-5267

I have used lipofectin (from Gibco). This may be dependent on the plasmid
to be transfected; but in my experience I got better results with
lipofectase. It is important this because Lipofectase can be easily
prepared. I have used this liposome for more than a year and I have had
very good transfection. The method that I use to prepare it is the one
described in

Rose et al. (1991) BioTechniques 10, 520-525

 The only that I can point out is that when I receive the PtdEtn, I
separate it in aliquots (enough for 1 ml of liposomes) in conic glass 15 ml
tubes, mix with DDAB and dry under N2. Then, I add 1 ml of water. In this
condition the tubes are stored in the refrigerator. When I need it, I
prepare the liposomes by sonication. I have used liposomes several months
old and have great transfection (measured by b-gal). I hope this help you.
Call me if you need other information.

carlos

______________________________________________________________________________
Carlos H. Pedemonte                     Telephone:  713-743-1211 (office)
Dept. of Pharmacology                               713-743-1228 (lab)
University of Houston                   Fax:        713-743-1229

Houston, TX 77204-5515                  e-mail: pedemonte at jetson.uh.edu
____________________________________________________________________________
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