Hello to all, glad to be part of the forums.
I'd like to know how important it is to know the DNA concentration of a given PCR product before purifying it? We are using a kit to purify the PCR product and then using the purified product in an RFLP assay. Several problems with these RFLP assays lead me to suspect that something is wrong with our purification process.
The PI says that finding the DNA concentration in our PCR product (via Qubit) is cost-prohibitive so we just go right to cleaning the product after amplification. Does anyone else go right to cleanup without verifying DNA concentrations, and if so how do you insure that you're not diluting the PCR product too much when eluting in the spin column?