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RNA extraction kits for small samples

rna isolation small sample

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6 replies to this topic

#1 Rute

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Posted 17 July 2012 - 05:44 AM

Dear all,

I use RNaeasy mini kit from Qiagen to extract RNA. However, when extracting RNA from really small samples (100x103 cells) the quality is terrible and the amount it's not that great.
I've heard of kits which were design for such small samples.

Can anyone advise me one?

Thanks.

Kind regards
Rute

#2 Curtis

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Posted 17 July 2012 - 09:21 PM

do you want to run RT-PCR? if so then just normalize your samples and go ahead. Don't worry about quality. How much is your concentration anyway?

You can alternatively use Trizol, but I can't promise a high quality with that.

#3 Adrian K

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Posted 22 July 2012 - 07:43 AM

I had seen there are some hybrid protocols which involved using both TRIZOL and RNeasy kit.
Hopefully this can help to solve both your yield and quality problem.
Here are some examples I found it from google:

http://people.oregon...easy hybrid.pdf
http://microarrays.h...croarrays12.pdf

Just my 2 cents.
Rgs,
Adrian
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#4 Rute

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Posted 23 July 2012 - 06:29 AM

I want to perform a microarray with the RNA thus the quality needs to be extremely good.

I saw the ones you suggested but htey won't do for me. The starting material that I have it's less then half that the one stated in the protocol.
I've heard of MagMAX™-96 Total RNA Isolation Kit but they also do not say the recomended startir material. Has any of you ever heard of this one?

https://products.inv.../product/AM1830

Edited by Rute, 23 July 2012 - 06:33 AM.


#5 Curtis

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Posted 23 July 2012 - 09:15 PM

I use Intron. Their kits usually work slightly better than Qiagen's.

#6 Trof

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Posted 01 August 2012 - 02:12 PM

In case of very small sample and the need for high amount and high purity RNA, I would consider some RNA amplification kits, if there is one suitable for microarrays.
You could get decent amount of RNA from small samples with Trizol, but you won't get microarray purity. If you use column to purify after that, you loose the RNA.
Only kits I heard of for small samples were those directly cell-to-Ct and such, where you amplify form kind of lysate and only for a special application, so not usable for microarray.

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#7 mdfenko

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Posted 03 August 2012 - 06:16 AM

these are for agilent gene expression microarrays:

quick amp labeling kit

and

low input quick amp labeling kit
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