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RT-PCR;Northern-blotting - internal standard (Apr/28/2004 )


I have two questions:

(1) I have done RT-PCR but the results are controry to my ideal, so I want to know how to quantify the template RNA so as to display the expression at the translation level.

(2) I am going to do Northern-blotting ,but I have no idea which company has better Northern-blotting kit available. If someone know something about these, would you like to share with me?

Thank you very much!


What do you mean by contraversy? Could you explain in more detail?


That is to say, the brightness of the bands in the gel should reflect the level of expression ,but my results were not compare to my observation of EGFP by naked eye.



To mark my probe, when I use 32P (dCTP from Amersham), I use the Rediprime kit form Amersham. It works OK!
And Genescreen membranes (PVDF or nylon) or also Nytran membranes Plus from Schleicher and Schuell.
But the prehibridation solution is not from a kit, I make it! Like the transfer buffer(SSC10X).

Good luck!
Bye! biggrin.gif



Thank you very much! I will have a try!