In vitro transcription ......

[Martin D. Leach]

In article <3j49va$hro at netnews.ntu.edu.tw>, r3211001 at cc.ntu.edu.tw
(r3211001) wrote:

> Dear netter,
> 
>     I want to find out the cis-regulatory elements of an oocyte-specifically 
> expressed gene. Because of some technique problems, I cannot transfect the 
> reporter constructs into the living cells. My advisor told me to use 'in 
> vitro transcription assay' in cell extracts. I've tried several times, 
> but it did not work.
> 
How about injecting supercoiled reporter constructs into the
oocytes...e.g. lacZ and assaying them...

alternatively...use a luciferase construct...and see if the oocytes glow
in the dark (scintillation counter with attached single photon monitor
would be a good choice for using a whole vial of oocytes) upon addition of
Lucifierin (K+ salt) - 5mM ? correct me.

Martin

-BTW I am not only the luciferase president...i am also a client..-

-- 
.....          Martin Leach                Email:leach at bu.edu 
   _|____      Dept. of Pharmacology       Phone: (617) 638-5323        
   / o  /      Boston Univ. School of Med. Fax:   (617) 638-4329         
 _/  |-/__==/  80 E. Concord St. (L603)
(BULLDOZER) \_ Boston MA 02118            "Not the old underpants 
               USA                                on your
                                             head.....WIBBLE"
My homepage: http://155.41.115.114/Leach.html
Biotech company listing: http://155.41.115.114/jumper/biolcomm.html

p.s. try BioMOO  -virtual biology on the internet..
telnet bioinfo.weizmann.ac.il 8888