Identifying an Unknown Sample Via Dna Fingerprinting

Identifying a bacteriophage used to treat MRSA Staphylococcus aureus via RFLP DNA Fingerprinting

Introduction

Restriction fragment length polymorphism (RFLP) DNA fingerprinting is a system that is used to cut genomic DNA into smaller segments which can then be separated and viewed under an ultraviolet light (Wang et al. 2016). In this experiment three known sample bacteriophages: X (ϕTCH60), Y (SAP-26) and Z (ϕSH19), and one unknown sample bacteriophage (E), will undergo RFLP DNA fingerprinting against a ladder (L) so that their separated segments can be seen and compared to each other (Li et al. 2010). The purpose of this is to find out which of the known samples the unknown sample matches up to. This will enable possible advancement in the treatment of antibiotic resistant golden staph (MRSA Staphylococcus aureus), a microflora which can be found on skin and nasal tissues of humans which can potentially cause major skin irritations (Arabi and Samuel. 2015). The technique of RFLP DNA fingerprinting has already been used in assisting scientists determine where unknown samples have come from when comparing with known samples and so the result of this experiment should have similar, trustworthy outcomes (Rea et al. 2009).

Aim

To determine which sample bacteriophage (X, Y or Z) has the same genomic DNA sequence as the unknown sample bacteriophage (E) through the use of RFLP DNA fingerprinting.

Material and Methods

Method and materials used can be found in the “Molecular & Cell Biology Practical Manual Semester II, 2016” (RSBANU, 2016). There were no deviations from this original method and apparatus. First the restriction enzyme and buffer were mixed with the four samples. Then the agarose gel was prepared and the four samples were placed separately into the wells of the gel to be run. The end result of this was then observed under UV light to be viewed.

Results

Figure 1. RFLP DNA Fingerprinting using “Restriction Enzyme”.

[pic 1][pic 2]

Summary

The bands from the RFLP can be clearly seen in the results. The DNA has separated well. The bands formed by X look like the bands formed by E.

Discussion

From figure 1 it can be observed that the bacteriophage X is most likely the same as the unknown bacteriophage E. However, their lines do not completely line up together and the experiment could be done once again with a more precise methodology of filling the wells to ensure that they are in fact the same bacteriophage. Compared to a similar experiment which compared and classified like bacteriophages of Pseudomonas aeruginosa, the standard of the results were below par. However the results did follow a similar method in determining which separated DNA segment sets were similar to each other (Li et al. 2010). This experiment managed to successfully discern the bacteriophage that matched an unknown sample. Therefore the same methodology can be followed to assist in other endeavours such as quality testing vegetables (testing the DNA of an unknown sample then comparing it to samples from where it claims to come from). In conclusion the identity of the bacteriophage E was found to be bacteriophage X, and so the advancement in treatment of MRSA Staphylococcus can be continued.