I'm aiming to attempt the opposite of what many on this forum are trying. I'm seeking to perform Trizol extraction with the implicit aim to purify RNA species that are longer than small RNA (miRNA, snRNA etc). I'm particularly interested in removing the miRNA component.
Trizol extraction by itself does a good job of being bad at co-purifying small RNA, but some small RNAs still make it through. Would anybody know a method to reduce the extraction of small RNAs without affecting the isolation of long RNA species?
Firstly, i am working since two year with miRNAs and I had the experience to tell you that Qiazol methode of extraction is not affecting the miRNAs population at all and you still able to detect it in your sample.
My advice in your quetion would be to go for Qiagen Kit called RNAeasy which include a sepration step in which a column is used that only trap the RNA that is longer than 200 nt but other small RNAs is just washed a way with the ethanole and RWT buffer.
Hi Sebastian
I agree about the use of miRVana. In fact the extraction kit comes with a protocol to extract total RNA and then separate the small species from the bigger ones. I used it with no problems from cells in culture.
Best,. Paco
Thanks all for the answers. I've used the mirVana kit before, but without the miRNA enrichment step. I was unaware that you simultaneously could obtain a fraction of >200 nt RNA.
However, I've only the RNeasy kit, so I'll give that a try.
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