Check the mRNA Quality - (Jul/12/2008 )

hi everybody
i want the method for check the mRNA quality for use constructe cDNA library, the kit use mRNA 1-5 ug.i should check it by spectrophotometer, electrophoresis or orther method ?
help me pls.
thank everybody

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Electrophoresis, but nowadays there are gadgets you can use.
Check these links:
http://search.vadlo.com/b/q?keys=RNA+quali...mp;sn=158621799
..

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i prefer using nanodrop... faster and easier.
also, you don't need to sacrifice too much sample (compare to running gels or spectro).

but if you decided to use 1 method, stick to that method/machine... to get a more consistent reading throughout your experiment.

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the nanodrop spectrophotometer can only get to know the concentration of sample, a260/a280, and a260/a230
i use agilent 2100 bioanalyzer to check my mRNA quality.. its much more efficient than run the electrophoresis gel. the common electrophoresis gel (Etbr) require around 1ug to clearly detect the sample. but using the bioanalyzer, the quality mRNA require can be from 50-500ng... only small quatity is require, so wont waste a lot sample.

compare to common electrophoresis gel, it also check the quality of ribosomal contamination in the sample.
to construct cDNA library, high purity of mRNA is require.
if the sample contain more than 5% ribosomal RNA, it might affect ur quality cDNA later..

hope this detail will be useful to u..
take care...

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yup. Bioanalyzer definitely give more information about your RNA.
If your lab has the budget for Bioanalyzer, go for bioanalyzer.
you'll need to run your sample through gel capillary using a chip. the principle is like running on a gel. you'll later get a profile of RNA band and also quality of RNA.
how ever, you need 12 samples for running i chip, or you can share with somebody else. also, chips are not cheap too.

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But if your lab or dept. dont have an agilen bioanalizer just run a denaturing agarose gel (check ambion webpage for the recipe).

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