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miRNA and TRIZOL


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7 replies to this topic

#1 borriello.87

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Posted 18 October 2009 - 03:17 PM

Hi everyone,
is it possible to extract also miRNA using TRIZOL protocol?

Thanks a lot!

#2 bob1

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Posted 18 October 2009 - 03:25 PM

Yes, just be sure that you don't use a column kit to clean it up as most don't capture miRNA.

#3 borriello.87

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Posted 19 October 2009 - 08:10 AM

Hi bob1, thank you for your answer!
How can I see miRNA in my sample? Can I see them with agarose gel or a Bioanalyzer is required?

Thanks again!

#4 EmilyG

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Posted 19 October 2009 - 12:00 PM

Hello

I'm afraid miRNAs are too small to be seen on an agarose gel and a Bioanalyzer will be needed. I've been using a pico chip but apparently you can get specialised small RNA chips to look at your miRNAs in more detail.

Hope that helps
BW
Emily

#5 Telomerase

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Posted 21 October 2009 - 03:27 AM

You can rt-pcr them with a kit, that contains specifically designed primers, with overhangs. This way you'll get the band on gel, of course. That is, if you're looking for a particular miRNA, not the whole bunch.
"Beware the power of a PhD student" - scolix

#6 gogreen

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Posted 21 October 2009 - 04:19 AM

you can use the bioanalyzer small RNA kit to check the miRNA. It gives the % of miRNA present in the sample. Its sometimes a bit confusing as it would show values ranging 13-80% of miRNA in your sample. So I always run a reference sample which gave me good miRNA microarray results before and while running the first chip, check with various dilutions of the RNA so that you can decide on the concentration which is optimal.

Edited by gogreen, 21 October 2009 - 04:21 AM.


#7 gogreen

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Posted 21 October 2009 - 04:20 AM

Hi everyone,
is it possible to extract also miRNA using TRIZOL protocol?

Thanks a lot!


Trizol gets most of the miRNA, but the Ambion mirVana kit used to give better results for me for the microarrays

#8 borriello.87

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Posted 30 October 2009 - 09:50 AM

Thank you, your answers have been very useful!




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