When I do RT-PCR, I use cDNA at a ratio of 1:100.18S value is good, but beta-actin ct value is 35.It also gives abnormal values in genes I use other than housekeeping.Can I run the cDNA in the gel? How can I control this?
Harsheema Ottappilakkil
Hi Meryem, have you checked your RNA for degradation? You can use a small aliquot of the total RNA and do an AGE to confirm the quality of your sample. You can also use a bioanalyzer for the same. Also, performing a standard curve analysis will help you determine the concentration of your template (1:100 might be too high) for further optimization of the PCR conditions.
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