Home-Made TA cloning

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>
>> I've been trying to think of a way to make my own TA cloning kit.  
>> 
>> There doesn't seem to be any available restriction enzymes that will 
>> produce the proper ends for TA cloning (although I'm sure Invitrogen 
must 
>> have one in their bag of tricks).


I am sure that Invitrogen is probably making their TA vector by first 
linearizing the polylinker with EcoR1 and then ligating an EcoR1 linker 
containing a T overhang.

		eg:    EcoR1 side
        			     5'	AATTCGGCTT  3'
					    GCCGA

Such a linker would be easy to make and fairly cheap since it is so 
short.  Unfortunately, I can't say that this method works as well in the 
lab as it does on paper because I am lazy and just buy the kit.

Christopher D. Skory
Agricultural Research Service
USDA/NCAUR
Peoria, IL