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miRNA yield from human urine

miRNA urine circulating miRNAs rna quality microRNA

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#1 itsron

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Posted 19 December 2014 - 08:28 PM

I recently isolated miRNA from human urine using Quigen miRNeasy serum/plasma kit (which works with urine as well) and quantified using NanoDrop. Even from freshly isolated sample, I am getting a horrible 260/280 ratios like 1.5 and 1.6. Concentration is around 8-30 ng/ul

 

My feeling is that the choice of Nanodrop is not right as miRNAs are only present in lower amounts in urine and these levels may not be detected accurately by NanoDrop. I can not imagine another reason for such a low ratio (I am pretty sure I did the experiment right).

 

Has anybody here worked with urinary miRNAs? I would like to know how much is the concentration range you are getting after isolation, and how to you quantify/qualify miRNAs?( I guess it would be Bioanalyzer).

 

Thanks mates!

 


Edited by itsron, 19 December 2014 - 08:30 PM.


#2 Curtis

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Posted 21 December 2014 - 04:08 AM

It could be the kit. Qiagen is one of the top kit producers, but I have got better concentrations from other kits made by other companies too. Don't want to mention any names here.



#3 itsron

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Posted 21 December 2014 - 09:12 PM

It could be the kit. Qiagen is one of the top kit producers, but I have got better concentrations from other kits made by other companies too. Don't want to mention any names here.

So which one do you suggest? Otherwise it is not going to help me!







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