Hi, everyone
I am extracting a total RNA from rat brain samples for real-time PCR studies, but unfortunately, there is DNA contamination in gel electrophoresis. I tried to remove it and I failed.
By the way, I prepare to don't use DNAse treatment. (maybe the last option)
which part may cause this problem you think?
Hi Solmaz
First: DNase treatment is an important criteria based on MIQE guideline for qPCR (Bustin et al, 2009). Perhaps you are considering the loos of RNA integrity during DNase treatment. I would recommend including commercially available RNase inhibitor during DNase treatment.
Second: If you need more hints, you may describe your RNA extraction procedure.
Good luck
John V. Stokes
Dear John,
Thank you for your answer.
The problem is DANse decrease my RNA concentration too, I prefer don't use it.
or maybe the last option.
Ali Javadmanesh
Dear Ali
Thank you for your response.
I will take this into consideration.
Hi Solmaz
I have been using this kit since 3 years ago. Reagent RL has a very short lifetime. Your kit might be old!
The good thing about brain tissue is that it has a very low RNase activity so don't be afraid to use DNase treatment.
Good luck
Ali Javadmanesh
Dear Ali,
I prepare a new kit, I don't think so. I will try DNase I treatment.
Thanks for your advice anyway.
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