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Use of the DCode System to Detect the Food-Borne Bacterial Pathogen Listeria monocytogenes

nts from different organisms to the same location on the gel, as displayed in Figure 1, does not necessarily imply that the fragments share the same sequence. Using a different primer pair (341F with GC-clamp and 534 R), we produced fragments from another region of the 16S rDNA from L. innocua and L. seeligeri, which differ in sequence by 2 bases (Figure 3). In this case however, no difference in the migration rate and final gel positions of the respective DGGE bands was observed (Figure 4). Co-migration may be explained, in this instance, in that the base differences did not change the %G+C of the melting domain in the fragments.

The PCR fragments generated from the DNA of L. innocua and L. seeligeri, using primer pair 1055F and 1406R with GC-clamp, also migrated to the same location in the gel (Figure 1). Examination of the base sequences of the fragments from these two species reveals sequence homology where base assignments have been made (Figure 2). Since base assignments have yet to be made at 9 locations on the fragment, whether the co-migrating fragments possess sequence homology or the same G+C content remains to be determined.

As was the case with primer pair 1055F and 1406R with GCclamp, the gene fragment obtained from L. monocytogenes with primer pair 341F with GC-clamp and 534 R yielded a DGGE band that was distinguishable from the bands obtained from the other two Listeria spp. (Figure 4). The base sequence of the fragment from L. monocytogenes differed from that of L. innocua by no more than 2 bases and no less than 1 base, and from that of L. seeligeri by no more than 1 base (Figure 3). The difference in migration rate of the band from L. monocytogenes compared to t
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