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.
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
Best of luck,
The University of Calgary
lmjnutte at acs.ucalgary.ca
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