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miRNA isolation from serum - (Mar/17/2010 )

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Hi comp3v,

You can email info@nanodrop.com and attach your file, or you can use the form in the "Contact Technical Support" (you can't attach data, but someone will get back to you and tell you what address to send it to). Your spectra does look terrible, but I am pretty sure that our tech support team will be able to help you.

Cheers.

-Limey-

Limey on May 13 2010, 03:50 PM said:

You can email info@nanodrop.com and attach your file, or you can use the form in the "Contact Technical Support" (you can't attach data, but someone will get back to you and tell you what address to send it to). Your spectra does look terrible, but I am pretty sure that our tech support team will be able to help you.
ah, I just thought that you are one of tech specialists and can comment the spectra by yourself. Ok, I've sent to them - hope to have any help.
(sorry people for this off-topic)

back to topic - question for those who used miRVana kit. After final elution (I do as written in manual: heat solution to 95 and apply 100 mkl to the column) I normally put my RNA samples to -80. When I thawed it back, I've noticed that solution is not completely clear. I made short high-speed centrifugation and got quite appreciable white pellet. This was reproducible for many samples. Did anybody have the same thing? (just try to centrifuge your sample)

-comp3v-

Man truth is your spectra are really mad !

About the precipitate, it's probably contaminents from plasma, like lipid or so. After freezing it may precipitate, I guess.
for the last step, the elution, try to apply 2 times 50ÁL. You should have a better recovery.

I realy don't trust the Nanodrop on such samples, but I guess we have to do with it...
I am going to run microarray, we'll see !

-Sero-

Dear Limey,

 

why do companies like Qiagen post on their FAQ that Nanodrop is not suitable for miRNA concentration measurememnt if it is not the case: https://www.qiagen.com/de/resources/faq?id=cc3d10b7-2d92-4fa3-91fd-a1f4e9999f13&lang=en ?

 

did anyone reach an answer to this? or an alternative?

 

Best,

Moe

-Moemedawar-

Dose the topic go to the problem on Nanodrop~biggrin.png

There is no problem for me to use Nanodrop to measurement RNA. But I did not find an item in Nanodrop to analysis miRNA.

It is possible the concentration of your RNA sample is too low, then the 260/280 ratio could goes down, too.

For me, I use Trizol, if my RNA sample concentration lower than 80ng, 260/280 ratio could be about 1.6.

So, I think the first thing to solve the problem is consider how to improve the concentration of your sample. 

-Andrea Fortina-
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