>>... I assumed that you were simply looking for positive or negative
plaques. I am also assuming that these are lambda phage and you have infected E. coli for your
> plaque lifts.
Yes
Are you getting light all across the entire membrane? or are
> you only getting light where there are plaques? Is the light amount
> proportional to the size of the plaques?
The plaques are well defined on film but the control plaques (those with
no secondary ab conjugate) are just as well defined as those with
secondary antibody conjugate. I think that exposure time is the problem.
Film results show higher plaque density than colorimetric methods as
well.
> manner. (As an aside comment, I and two other independent researchers in
> our Dept. tried chemiluminescent procedures and were never happy with the
> inconsistent results.) A final set of questions: how does the colorimetric
> method compare to the chemiluminescent results? Does the coloimetric method
> work and the chemiluminescent not work or vice versa?
I am getting positive results from both methods but chemiluminescence is
so sensitive that I think it may be impractical.
Thanks for your help.
--P. Murphy
