I get multiple bands on qPCR, and multiple peaks in the melt curve. this question is an attempt to understand whether there is DNA contamination even after fractionation and Trizol RNA isolation.
Hi, Shweta Warrier
I have no answer for you but your question has intrigued me and made me decide to write down a few lines.
1 - Probably yes. All different techniques bring us "closer to zero" contamination but not an absolute zero.
2 - In the specific case of ultracentrifugation, every single structure/molecule in the tube will be forced to go down ... (You didn't give an idea of how many steps before ultracentrifugation you may have performed), So, I believe "yes", you have lots of different contaminants, although a higher yield of your target polysomes.
3 - If you have done specific primers for the sequences you're looking for at the polysomes (and these sequences are not shared by other substructures or from the nucleus), your results have not been disturbed by the external DNA (not from polysomes).
4 - Finally, I think you can perform a qPCR/PCR for an specific, usually, highly abundant DNA sequence (Not present in polysomes) to check it and find out If you have DNA in your samples or not.
Hope it helps.
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