Reverse Northern

Paul N Hengen pnh at ncifcrf.gov
Tue Sep 24 11:27:09 EST 1996


Gene Huh (gshuh at garnet.berkeley.edu) wrote:

| What I'm missing is exactly what a RN (reverse northern) is supposed to be,
| and who started this wacky terminology? Is it blotted cDNA probed with labeled
| RNA? What is Eberwein technology?

> My concept of an RN (others may differ) is that of dot/slot blotted,
> cloned cDNA sequences that are then probed with radiolabeled cDNA.  The
> probe cDNA is reverse transcribed from total or poly(A) RNA.  Hence the
> signal on a slot blot reflects the relative abundance of the mRNA
> represented by the cDNA in that particular slot.  Yes, it's a little
> wacky, which was my point to begin with :).  "Eberwein technology" was my
> shorthand for a variant of "RN" where the cDNA used as a probe is first
> converted to RNA (via T7 transcription) before probing.  

I agree with you that the term "expression profiling" seems better in this
context, but it is really a general term and could be mistaken for many
things. Does anyone have the exact reference from J. Eberwein, or the one
mentioned that was published in Nucleic Acids Research? Does anyone know
of a better primary reference for the method?

Technically speaking, is there any advantage to making the RNA probe
through T7 RNA polymerase first?

-Paul.

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