A GOOD KIT FOR PCR CLONING

[David Micklem]

In article <reedc-2901952101470001 at ip043167.iac.net>, reedc at iac.net (Reed
R. Corderman) wrote:

> In article <Pine.ULT.3.91a.950126165357.24016A-100000 at kepler.unh.edu>,
> xz at KEPLER.UNH.EDU (Xin-Hua Zhou) wrote:
> 
> >  I am looking for a good kit for PCR cloning. Can you give me some ideas 
> > , based on your EXPERENCE?
> 
> The best so far for me is the TA cloning kit from Invitrogen.  With a 25
> microliter PCR I use 2 to 4 microliters directly in 25 microliters of
> competent cells. Follow their transformation protocol, plate the entire
> transformation on two plates and voila!- Bunches of white colonies and a
> few blue ones.

Er.  Maybe I'm missing something here.  Don't you need a ligation in there
somewhere between the PCR and the transformation?  If not, I'd be very
interested to hear the protocol!

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D.R.Micklem,
Wellcome/CRC Institute,       Time flies like an arrow...
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Cambridge CB2 1QR             Fruit flies like a banana.
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Tel: [+44] (0)223 334129      Email:drm21 at mole.bio.cam.ac.uk
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