In: Biology
DNA sequence may be used to identify different species. Assuming that you can use PCR to generate products for sequencing, describe in detail an experiment that will allow you to determine which strain of Salmonella has caused the more recent outbreak.
according to recent studie and nomenclature, the genus Salmonella contains only two species, Salmonella enterica and Salmonella bongori. S. enterica is subdivided into six subspecies designated as follows: enterica (I), salamae (II), arizonae (IIIa), diarizonae (IIIb), houtenae (IV), and indica (VI), while S. bongori is not divided into subspecies. In PCR we can amplify a oligonunleotide/polynucleotide sequence using a primer, that is complementary of that sequence.
1. 1st we can determine different conserved sequences in different Salmonella.
2. Then we can make probes complementary to those conserved sequence and will amplify the no. of these probes by PCR.
3. when all probes are ready the, probes from each strain were tagged with different markers. It can be a molecule which shows radioactivity. or it can be a fluorescent molecule. when a specific probe will bind to a sequence in DND, it will show either florescence or radioactivity. This will prove the presence of that species of bacteria.
4. now bacterial cell ( suppose Salmonella bongori ) which is collected from infected individual, from that DNA is isolated and southern blotting is performed. When the DNA from test sample is exposed to several different probes(specific for different species and subspecies), only probe specific for Salmonella bongori will bind to it. and will show radioactivity. By this method we can test each sample. and can quantify the no. of cases related to each strain during and outbreak