Determining quality of polyA+ RNA
Hofstra at aa.wl.com
Thu Nov 2 14:55:25 EST 1995
jan016 at lulu.acns.nwu.edu (Jeff Nemeth) wrote:
>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.
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.
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