DDRT-PCR Northerns

Mayumi Yagi myagi at u.washington.edu
Thu Jun 29 12:16:16 EST 1995


Does it have to be a Northern?  Could you get away with either cloning 
out the relevant cDNAs and/or just doing RNase protection assays?  I'm 
thinking that the RNAs you're looking at are relatively rare, and RNase 
protection is supposed to be severalfold more senstive than a Northern.  
Of course, it won't give you size info, but if all you want to show is 
that it is indeed differentially expressed, maybe this will let you 
finish.  I've had good luck with the RPAII kit (and MEGAscript) from 
Ambion (the usual disclaimers in this case).  Good luck!

On 25 Jun 1995, Shahram Mori wrote:

> Dear Molbionetters and sufferers of DDRT,
> I have worked long and hard and managed to get all the stages of DDRT to 
> work. I am isolating bands that to look differentially expressed and 
> repressed, however the northerns don't work. How many people have 
> observed this out there? What did you do? Strauss et al in PCR Methods 
> and Applications said that they ahd given up doing Northerns without 
> giving a clear reason why. 
> I wanna graduate and this god-damn northern doesn't let me.... :,,(
> Yes, I wanna leave New Mexico (nice place though) and go to San Diego. 
> But I won't be able to now... I hate DDRT. Liang; are you hearing this??????
> Cheers
> 
> --
> 				_/\_
> 			       _\  /_
> 			       \_  _/
> 				 ||
> 			C   A   N   A   D   A
> DNA$DNA$DNA$DNA$DNA$DNA$DNA$DNA$DNA$DNA$DNA$DNA$DNA$DNA$DNA$DNA
> Shahram Mori
> Program in Molecular Biology
> Department of Chemistry and Biochemistry Box 3C
> New Mexico State University
> Las Cruces NM
> 88003
> RNA$RNA$RNA$RNA$RNA$RNA$RNA$RNA$RNA$RNA$RNA$RNA$RNA$RNA$RNA$RNA		
> 
> 
> 
> 



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