Determining quality of polyA+ RNA

Angela Hofstra Hofstra at aa.wl.com
Thu Nov 2 14:55:25 EST 1995


jan016 at lulu.acns.nwu.edu (Jeff Nemeth) wrote:
>Marc,
>Sorry I don't have an answer for you, but a question.  I have been 
>isolating mRNA from small amounts of tissue, and always have a very hard 
>time quantitating the RNA.  I have too little to measure on a 
>spectrophotometer, and DNA dipsticks are awful.  Is it possible to 
>quantitate by running mRNA on a gel?  How much do you need to run to be 
>able to see 18 and 28s bands?  Agarose or acrylamide gel?  
>
>Any advice you may have would be greatly appreciated.  I'm desperate.
> Jeff N.

Jeff,


What about serial dilutions of your mRNA on a dot or slot blot, with a 

pure poly(A) standard curve and a poly(T) probe.  Farrell in "RNA 

Methodologies suggests doing this to normalize total RNA for poly(A) 

content.  It works with only 500 ng of total RNA so I would think you 

wouldn't need to sacrifice much of your mRNA.

Angela




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