Wretched dinucleotide repeats

Dr. Peter Gegenheimer pgegen at kuhub.cc.ukans.edu
Tue Jun 4 16:16:52 EST 1996


In <NEWTNews.833485390.12682.mullen at SMALL3.jsei.ucla.edu>, mullen at SMALL3.jsei.ucla.edu writes:
>In article <4okhac$ljk at sol.sun.csd.unb.ca>, <sgriffit at rpc.unb.ca> writes:
>> 
>>  We're using selected bovine microsattelites as tools in forensic 
>> analysis of moose and white tailed deer material to catch poachers.As 
>> with any dinucleotide repeat system the real allele holds court to an 
>> xtra band(s?) as a result of T addition by Taq ,as well as the lower 
>> molecular weight stutter bands that I'm lead to believe result from the 
>> collapse or shift of repeat units.I've been dickering around with T4 
>> polymerase which is supposed to nip the T overhangs off the amplified 
>> product, but ,so far,the enzyme and 5X buffer supplied by Gibco don't 
>> appear to have much effect. Has anyone had any success in reducing the 
>> appearance of these additions or shortened products? Is there any way of 
>> preventing the stutter effect?
>>        Thanks for checking in 
>>                                  STEVEg

Is is not possible to use one of the many proof-reading DNA polymerases, such as VENT, Pfu, etc? They certainly won't add extra bases, though whether they'll produce an amplification p[attern similar to Taq is a matter for trial & error.

P. Gegenehiemr/U. Kansas Biochemistry/Lawrence KS 66045-2106



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