Need advice on dT tailing of blunt end cut pBluescript

Zhiyong Zhao zxz4 at PSUVM.PSU.EDU
Wed Apr 16 11:30:35 EST 1997


> Need advice on dT tailing of blunt end cut pBluescript
> 
> Iain Shaw (iain.shaw at bbsrc.ac.uk)
> Wed, 16 Apr 1997 09:38:17 +0100
> 
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> ----------------------------------------------------------------------
> 
>
> 
> Dear All,
> 
> I am currently cloning PCR reagents into invitrogen TA kit, quite
> succesfully but at quite a high cost. What I would like to do is make
> my
> own by cutting pBluescript and add a single T overhang. I have a
> T-tailing kit from Boehringer (DNA Tailing Kit), but you do not appear
> to
> be able to limit the amount of Ts that are added. Does anyone know a
> way
> of adding a single T this way, or has anyone any suggestions on how
> else
> to do this.
> 
> Thanks in advance for any help you can give.
> 
> Iain Shaw
> ----------------------------------------------------------------------


Polish the ends of PCR products with Klenow. After PCR is done, add Klenow 
into the tubes. After the ends are filled, purify the PCR products.

Good luck.


Zhiyong Zhao



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