Question: Klebsiella pneumoniae is a Gram-negative bacterium that causes destructive changes to human lungs; inflammation and hemorrhage. K. pneumonia is a member of B-lactamase bacteria which have mechanisms to increase the resistance against wide range of antibiotics. ampC gene is one of genes are responsible to produce these lactamases.
In order to diagnose Klebsiella species collected from hospitals, a researcher targeted the ampC gene to classify the collected isolates. Two primers were designed to amplify this gene
Forward: ACCTATACGGCAGGCGG;
Reverse: GCCCGTTTTATGCACCCA.
The amplification conditions were optimized using a positive control. On the experiment level, the primers were able to amplify a nice amplicon. However, the sequencing results showed a false product.
1. Can you explain this conflict and suggest the source of error?
2. Can you tell how to design the experiment properly? Please show your answer in steps.
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