TA ligation - molecular ecology

Andrew O'Neill oneill at florey.biosci.uq.edu.au
Fri Apr 3 00:58:26 EST 1998

I am experiencing difficulty in ligating purified PCR amplified, mixed community
16SrDNA from a sediment sample.  The kit is manufactured by Promega and all 
controls, including ligation control are working according to specs.  However, my 
sample does not ligate into the pGEM-T vector.  Any tips, experience with this 
problem? I have also tried excising the 1.5kb band from an agarose gel and 
purifying, and an A-tailing reaction post-PCR.  

I would really appreciate any advice sent to my mailing address, as e-mail access is 
more readily available than internet.

Thanks in advance,

Andrew O'Neill 

Andrew O'Neill B.Sc. (Hons) PhD student
Centre for Bacterial Diversity and Identification
University of Qld, St Lucia, Qld, 4067
email: oneill at florey.biosci.uq.edu.au
ph: +61-7-3365-3211
fax: +61-7-3365-1566

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