I am working on determining the gene splicing and protein level in gene ko mice compared to wt ones. Since some of my results show inconsistencies, I'm thinking about pooling my samples to see if I can get a better result. Because I need to normalize each of my results and combine them, I think it would be easier if I combine them at the first point.
Have anyone used pooled samples for experiments like electrophoresis/western/flow? Any recommendation on the sample number?
If, without pooling your samples, you do see inconsistencies in the results, then pooling them will show even more inconsistencies because you introduce additional variables by pooling. I would iron out the procedure and try to come up with consistent results before doing any sample pooling. Also, do you have positive and negative controls for your experiment? Having these controls will help a lot in troubleshooting your "inconsistent" results. HTH.
Thanks Vicente. And yes, I have controls in my experiments. Let me refine my problem: I'm working on alternative splicing, and some of my biological replicates show different degree of isoforms expression. For example, my sample 1 shows high level of isoform 1&2, sample 2 shows high level of isoform 1&3 but not 2, sample 3 shows moderate expression of 1&2 but not 3. In this case, I know there are three isoforms present, I just don't know how to combine these results together.