About: Pseudomonas frederiksbergenis is a gram-negative bacteria that was originally identified in Frederiksberg, Copenhagen, Denmark at a coal gasification site. This bacteria is rod shaped, and colonies are known to be a pale yellow. Pseudomonas is known to be a species that is important for soil, food and water decomposition.
Date Collected: 2/8/2017
Methods of isolation and characterization:
- P. frederiksbergenis was isolated from a specific soil sample at the Environmental Education Center (EEC) located in Farmville, VA.
- The soil sample was taken directly from Buffalo Creek, a small creek that flows into the Appomattox river.
- Once we collected our sample we plated it onto three agar plates: direct sample, 1/10 and 1/100 dilution.
- We observed growth at 24hrs and 48hrs and selected a specific colony to analyze that was not overlapping other colonies.
- We ran DNA isolation and purification through Nano Drop to ensure we had a pure sample to then proceed to PCR amplification of 16s rRNA.
- After isolating 16s rRNA of our colony we ran gel electrophoresis against MSp1 to ensure we isolated 16s rRNA to send off for sequencing.
- Sadly we were unable to draw any conclusions from our gel due to clarity issues, never the less we went ahead and sent off our sample off to be sequenced.
- Once sequenced, we than took our sequence and ran it through BLAST analysis which gave us a 99% match
- Using our data analysis from BLAST as well as the color, form, and size of our colony we identified our sample to be P. frederiksbergenis.
Results:
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Open in new tab Figure 1. Chromatogram of our Buffalo Creek soil sample, which we identified as P. frederiksbergenis.
Figure 2. Buffalo Creek soil sample and location in which we isolated P. frederiksbergenis.
Figure 3. Our colony of P. frederiksbergenis under a microscope. This photo highlights the pale yellow color that we used to further support our classification.
Figure 4. The 1/100 dilution plate of the Buffalo Creek soil shows the specific colony we isolated P. frederiksbergenis.
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