Reverse Northern Blot?

szcooley at chip.ucdavis.edu szcooley at chip.ucdavis.edu
Wed Sep 28 11:08:00 EST 1994


Dr. YL Du (yd128 at albnyvms.bitnet) wrote:
: Hello:  I have a question for the Reverse Northern Blot.  Basically, I
: want to use reverse transcriptase , poly T oligo, dNTP, and 32p-dCTP to
: synthesize labeled cDNA.  Then using this as probe to hybridize with
: spotted plasmids on nylon membrane.  Is there a protocol for this?  Any
: experience?  All of suggestions are welcome.  Thanks.

I did this using the protocol which came with the RT (MMTV from BRL), but 
it did not produce very high specific activity. I have heard of a 
protocol whereby you make cold first strand then use any random primer 
labelling kit to label the second strand.


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