Restriction in a PCR tube

Duncan Clark Duncan at genesys.demon.co.uk
Tue Nov 2 16:20:08 EST 1993


In article <9311020154.AA18023 at cscgpo.anu.edu.au> Klaus.Matthaei at ANU.EDU.AU writes:

>>Hello netters!
>>
>>        Has somebody try to digest PCR product in PCR tube (I mean in PCR
>>reaction mixture) with different restriction enzymes? 
>>        
>

Unless you inactivate the Taq pol in the original PCR, when you cut with the 
RE (which isn't a problem) the polymerase will blunt your DNA. This is 'cos 
the enzyme is still active at 37C and dNTP's used in the PCR are still be 
present. 

Duncan  
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