Methylation detection??

Paul Rohde P.Rohde at mailbox.uq.edu.au
Sat Oct 4 05:07:17 EST 1997


Sorry for my ignorant responce, and I don't know if it is applicable
to you, but there is a chemical method that transforms the methylated
base from one type of base to another.

So you PCR half your sample and sequence it.
On the other sample, you treat it, PCR and sequence, then look
for the base change differences (transformed bases).

bye


woodlab at SUNGCG.USOUTHAL.EDU (Wood Lab) writes: > 
> 
> Hello out there,
> 	I am trying to determine if there is any methylation in the 
> bacterial system I am using.  So far, I have done the straight-forward 
> restriction digestions, using HinfII, etc.  No methylation detected.  I 
> have been reading about the bisulfate modification of the cytosines and 
> I'll probably give it a try.  My concern is that my critter is very A/T 
> rich and only about .75% of E. coli's Cytosines are methylated.  I am 
> afraid I'll miss it.  I have also noted that someone determined 1% of 
> the adenines of E. coli are methylated.  How did they determine that??  
> If I could check for methylation of adenine, that would be great.  
> 
> If any of you have information on this or any methods I haven't heard of, 
> I would greatly appreciate it.  Oh, Mass-spec-Mass-spec may be out of the 
> question, but I'll take info or references on that as well.
> 
> Thanks in advance,
> 
> Aimee Tucker
> 
> 
> Department of Microbiology
> University of South Alabama
> e-mail woodlab at sungcg.usouthal.edu




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