5 replies to this topic

[jac05021]

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

[pDNA]

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

[jac05021]

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

[jac05021]

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

[GHANBAR]

hello

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

THANK you

[pDNA]

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

Best,
p