Isolation of protein and RNA from RNeasy Minikits? - (May/06/2013 )

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

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

concerning the RNeasy columns ...you purify the RNA using the column and then you precipitate the flow through, like here:
http://www.biotechniques.com/multimedia/archive/00005/BTN_A_06401BM05_O_5037a.pdf

Precipitation is also described here.

Regards,
p

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

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

hello

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

THANK you

http://openwetware.org/wiki/RNA_extraction_using_trizol/tri

Best,
p