Amplification of E. coli uidA using PCR.
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The results of my PCR experiment on amplification of the portion of the E. coli uidA gene
The polymerized chain reaction is one of the fundamental procedures used in the
laboratory in the identification of bacteria on a molecular basis. The polymerized chain reaction
is so effective and highly accurate in the accountability of identification of a strain of bacteria
using the molecular base pairs in ac. Using the same method, an experiment was done in the
laboratory where the following results were obtained.
Results
After the amplification of the uidA gene from E.coli, I analyzed the sequence and come
into the conclusion that the part carried in 350bp specifically of glucuronidase gene uidA
presents a mutation. These findings were astonishing when I discovered that from the
experiment, the portions similar to the one I analyzed carried out the mutation characteristics. A
specific position mutated was in the bases where T had been altered to G.
When the PNA was not incuded known to amplify the products corresponding to uidA
genes were obtained. The bands were then detected from the strains. Thus the data concluded
that, the PNA used the inhibition mechanism in restricting the PCR amplification of the uidA
gene. The PCR clamping is only successful when the peptide nucleotides are included.
Furthermore, more details were obtained when the DNA primer and the peptide nucleic
acids coined as PNA reveals that, this nucleic acid has high thermal stability than the DNA
complex itself. In addition, the analyzation of the uidA gene was kept in the gel electrophoresis
of a 3% agarose gel, and the mixture was stained with the chemical liquid ethidium bromide.
This generates an image as shown below, extracted from the PowerPoint working out.
PCR Amplification of the uidA Gene of E. coli
M 1 2 3 4 5 6 7 8 9 10 11 12 13 14
500 bp
600 bp
500 bp M 15 16 17 18 19 20 21 22 23 24 25 26 27 28
600 bp
The data table for the samples that were used during the PCR analysis experiment for the
uidA.
BACTERIAL SEROTYPE NUMBER uidA gene Type of shigatoxin
SPECIES OF in multiplex toxin
STRAINS product
E. coli O157:H7 2 + Stx.stx2 +
O157:H7 2 + Stxl +
O157:H7 1 _ Stx 2 _
O28ac 1 _ Stx 2 +
O55 1 _ Stx2vp +
O139 1 _ +
O1 1 _ _
O2 2 _ _
O5 1 _ _
O78 1 _ _
s. urbana O30 1 _ _
s.ryphimuriu O4 1 _ _
S, enterifids O9 1 _ _
c.freundii 1 _ _
References
Molina, F., López-Acedo, E., Tabla, R., Roa, I., Gómez, A., & Rebollo, J. E. (2015).
Improved detection of Escherichia coli and coliform bacteria by multiplex
PCR. BMC biotechnology, 15(1), 1-9.
Taskin, B., Gozen, A. G., & Duran, M. (2011). Selective quantification of viable
Escherichia coli bacteria in biosolids by quantitative PCR with propidium
monoazide modification. Applied and environmental microbiology, 77(13), 4329-
4335.
Website,
https://www.google.com/search?q=a+properly+formatted+fi
Yoshitomi, K. J., Jinneman, K. C., & Weagant, S. D. (2006). Detection of Shiga toxin
genes stx1, stx2, and the+ 93 uidA mutation of E. coli O157: H7/H-using SYBR®
Green I in a real-time multiplex PCR. Molecular and cellular probes, 20(1), 31-41.
