attempt to amplify a product from intestinal cDNA, but this was successful only for the

primers that annealed to the untranslated sequences (Figure 5-21).


0)1234567


8 9


0)123456


1500 --)


Figure 5-20. Cloning of livUGTn.


A. Amplification of UGT gene with (lanes 1-6) and without UTR (lanes 7-9),
using various primer combinations and annealing temperatures. B. UTR insert
(lanes 1-3) and UGT insert (lanes 4-6) cloned into p-GEM T-Easy vector, total
size ~ 4500bp. 1kb ExactGene DNA ladder shown in first lane (0) of both
pictures.
A B


0 12 34 0


1234


1500 --)


Figure 5-21. Cloning of intUGTn.

 A. Amplification of UGT gene with (lane 3) and without UTR (lane 4); lanes 1
 and 2 show unsuccessful amplification with degenerate primers. B. UTR insert
 (lanes 1-4) cloned into p-GEM T-Easy vector, total size ~ 4500bp. 1kb ExactGene
 DNA ladder shown in first lane (0) of both pictures.