I am doing competitive quantitative RT-PCR where target DNA and standard are
amplified together. The standard is similar to target sequence except for a
small deletion. When I amplify each on its own , I have no problem and a
single product is observed , but when the two are amplified together, at
concentration that allow both to be amplified, I see a product amplified from
target, a product from standard, and a third band in between the two that I
can not relate to anything.
Has anyone came across this problem before ?
If yes, could you please advice on how to solve this problem.