genomic sequencing

Warren Gallin wgallin at gpu.srv.ualberta.ca
Sun Oct 19 10:04:42 EST 1997


In Article <tonygt19-1810972211110001 at ts007d40.den-co.concentric.net>,
tonygt19 at concentric.net (Anthony G. Gutierrez) wrote:
>In article <62597k$trc$1 at news.iastate.edu>, lmarek at iastate.edu (Laura F
>Marek) wrote:
>
>> Can anyone give me information about sequencing using genomic DNA as
>>  template and/or how to find any information about this topic? 
>>  A search of this group's archives yielded only that a few others
>>  have posted a similar question; no answers.  Thanks very much.
>> 
>> Laura Fredrick Marek
>> G313 Agronomy
>> Iowa State University
>> Ames IA 50011
>> lmarek at iastate.edu
>> 
>>
>It's probably not possible to directly sequence from genomic DNA since you
can't control the quantity of the stretch of DNA to be extended. You are
also likely to have multiple primer binding sites which will make your gel
look like hell. If you have sequencing primers for a targeted region why not
amplify that first, purify it and then sequence the amplicon? 
> 
>Tony Gutierrez
>Molecular Bio Lab
>USACHPPM, DSA-West  


Not correct.  Check out a paper by Church and Gilbert, PNAS, Some time in
the last 10-15 years.  They describe their method for direct sequencing of
genomic DNA.  Note: it involves a massive hybridization blot, not the more
standard sequencing technologies.

Warren Gallin
Department of Biological Sciences
University of Alberta
Edmonton,  Alberta     T6G 2E9
Canada
wgallin at gpu.srv.ualberta.ca



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