| | Re: no amplification or non specific band
I order to figure out what is wrong in your reactions, you need to be a little bit more detailed on
1. What organisms are you extraction from
2. How do you do reverse transcription
3. What primers are u using
4. What are the primer and DNA concentrations
5. What are the PCR conditions
In general, it is helpful to measure DNA concentration but that gives you little info about the Quality of your DNA. If the material you are extracting is old, then your DNA might not be the best for amplification. Hence you get some samples with good PCR bands and others just won't work. Also, if your DNA concentration is too high, you will get a smear rather than a nice band.
The same is true for the primer concentration (more than one band when using too much)
So, in order to really tell what went wrong, we need more info!!!