GFP reporter gene work

Malcolm Ogg rmhamso at ucl.ac.uk
Fri Jul 31 04:17:54 EST 1998


In search of help:

I am hoping to begin work in my research group using GFP variants as
reporter genes in promoter assays.

The basic aim will be to co-transfect mammalian cells with one variant
to normalise for transfection efficiency, and another variant as a
construct containing the promoter region under examination.

I would like to carry this out in 96 well plates, with detection on a
fluorescence microplate reader.

I would be extremely interested to hear from anybody with any experience
with this situation.  I would like to know what GFP variants (colour and
supplier) people have used for 'dual reporter' assays, and whether
sufficient expression is readily detectable in most mammalian cells in
96 well plates.

I be interested in any comments or criticisms (send direct to me rather
than posting on the list if appropriate) about plasmids from individual
manufacturers and also about which fluorescence plate readers to buy or
avoid.

Thank you for your patience and time
Yours sincerely

Malcolm S Ogg
Dr. Malcolm S. Ogg
Centre for Cardiovascular Genetics
Department of Medicine
UCLMS
The Rayne Institute
5 University Street
London
WC1E 6JJ
Tel: 0171 209 6977
Fax: 0171 209 6212
Email: rmhamso at ucl.ac.uk



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