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Just a question about the RNA purification procedure. - Did...

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Ujunwa C Okoye-Okafor Aug 2, 2018
protocol Protocol: Dot Blot Analysis of N6-methyladenosine RNA Modification Levels
Just a question about the RNA purification procedure.

- Did anyone attempt this on total RNA and if yes how did you extract it?
Many thanks
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Hao Yu Author Answered Aug 2, 2018

Temasek Life Sciences Laboratory and Department of Biological Sciences, National University of Singapore, Singapore

In our experiments, we used RNeasy plus mini kit (QIAGEN) to extract the total RNA. We also tried to this protocol on total RNA, it worked. However, please note that m6A is also found in rRNA, tRNA and snRNA.
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Yu Liu Answered Aug 2, 2018

UH

I have limited experience with this protocol. It seems that the mRNA quantity is very important. 20 ug mRNA perhaps is the lower limit for decent signals. I imagine that you would need 200 ug total RNA to have the same amount of signal. Why don't you try Trizol and let us know the results.
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