not being able to reproduce result - (Dec/15/2009 )
Hi,
i am facing one trouble in my RT PCR results of histone acetylation study. I did 10 of my antibody samples in one 96 well plate, and did IgG of those 10 samples in other , so antibody and IgG are in diffrent expts. and then incorporated the result. Result was interesting to us.
2nd time i did antibody and IgG samples (same samples as of 1st expt) together in one plate, The result did not match with 1st incorporated result. So i tried one more time (3rd time) doing antibody and IgG together in one plate, could reproduce the 2nd result . The result is no more interesting.
So my question is what might be the reason for not getting the 1st result? I know primer efficiency varies b\w expts, what might be the other reasons?
thanks,
epigenetics on Dec 15 2009, 07:31 AM said:
i am facing one trouble in my RT PCR results of histone acetylation study. I did 10 of my antibody samples in one 96 well plate, and did IgG of those 10 samples in other , so antibody and IgG are in diffrent expts. and then incorporated the result. Result was interesting to us.
2nd time i did antibody and IgG samples (same samples as of 1st expt) together in one plate, The result did not match with 1st incorporated result. So i tried one more time (3rd time) doing antibody and IgG together in one plate, could reproduce the 2nd result . The result is no more interesting.
So my question is what might be the reason for not getting the 1st result? I know primer efficiency varies b\w expts, what might be the other reasons?
thanks,
It depends, are your samples from the same origin?
Are you sure that they are in the exact same state as the samples that you used in your first experiment?