Anyone successfully doing RNAse protection Assays?

Lauryl Maiya J Nutter lmjnutte at acs5.acs.ucalgary.ca
Thu Feb 29 15:43:31 EST 1996


genes at unixg.ubc.ca wrote:
: I have started doing RNAse protection assays using an Ambion kit(RNAse 
: II). I can't seem to get it to work. The probe is 600 bp and is made 
: from a pGEM plasmid containing an insert. The target is made from a 
: longer sequence that is in another plasmid (probe was actually 
: subcloned from this longer sequnce). I am trying to titre the reaction 
: so as not to use up my total RNA sample that is limited. The control 
: for the kit works fine. Can anyone give me some pointers on making the 
: probe or the target sample and getting this to work. 

Craig,

When you say that it doesn't work, what do you mean?  Are you
getting a blank film?  If so, what isotope are you using to label
your probe?  If not, what do the bands look like?  Full length?
One or two smaller bands? or a bunch of smaller bands resembling
a ladder?  Perhaps if I have more details I can give you some
suggestions.  

Best of luck,

Lauryl Nutter
Biological Sciences
The University of Calgary
lmjnutte at acs.ucalgary.ca



More information about the Methods mailing list