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Isolation of protein and RNA from RNeasy Minikits?

rna western blotting protein extraction RNeasy

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5 replies to this topic

#1 jac05021

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Posted 06 May 2013 - 08:02 AM

Hey everyone,

So I'm trying to see if I can isolate protein and RNA from the RNeasy Minikit. I am working with about 7 mg of rat nasal epithelial tissues and therefore need an efficient way to get both RNA and proteins from the same small sample. I know they sell a DNA/RNA/protein kit from Qiagen, but it is very expensive. TRIzol is also not going to be an option because I have not had good luck isolating RNA from my samples with their methods (contamination issues and low RNA yields).

Any suggestions on how I can isolate both total protein and RNA from my rat samples using this RNeasy method (or one just as costly...or cheaper :-) ).

Thanks!

Joe

#2 pDNA

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Posted 06 May 2013 - 10:19 AM

do you tried this one (uses Trizol):
http://www.ncbi.nlm....pubmed/17489233

concerning the RNeasy columns ...you purify the RNA using the column and then you precipitate the flow through, like here:
http://www.biotechni...M05_O_5037a.pdf

Precipitation is also described here.

Regards,
p

#3 jac05021

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Posted 06 May 2013 - 12:24 PM

Thanks! The RNeasy column protocol will be the one i try next! thanks

#4 jac05021

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Posted 10 May 2013 - 12:56 PM

by the way, this protocol worked beautifully. I did make some editions though.

I find it better to add 1.5 volumes of IPA to the 1st flow through from the RNeasy column (ie from the 70% EtOH addition step). I then continue with the protocol verbatum. It saves on having to wait a day of letting your tubes sit at -20C. I was able to get RNA and whole cell lysate protein samples from the same 4-5mg chunk of nasal tissue!

Thanks again.

Joe

#5 GHANBAR

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Posted 11 May 2013 - 01:22 AM

hello

I need protocol RNA extraction that cheap and have good qualify.

THANK you

#6 pDNA

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Posted 12 May 2013 - 08:55 AM

http://openwetware.o...sing_trizol/tri

Best,
p





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