Running head: DNA Fingerprinting 1

DNA Fingerprinting

Allisa McAleese

Breckinridge School of Nursing

Dr. Lok

GE 257
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DNA Fingerprinting

1. What kind of patient samples are used for the purpose of identifying possible pathogens?

There are many types of samples, usually types of bodily fluids, which can be used to

identify possible pathogens. For the pathogens identified in this virtual lab, fluid from

Lymph Node (Bartonella henselae), stool (Escherichia coli or Yersinia enterocolitica),

urine (Pseudomonas aeruginosa), blood (Salmonella typhimurium), and sputum (Yersinia

pestis) were used (Black, 2012).

2. What does PCR do, how does it work, and why is it useful?

PCR stands for polymerase chain reaction, and is a scientific technique used in molecular

biology to amplify a single or few pieces of DNA across several orders of magnitude.

Doing this generates thousands to millions of copies of a particular DNA sequence. It's an

inexpensive technique that can make segments of DNA. PCR can target specifics parts of

DNA (Black, 2012).

3. How do you separate the desired DNA from all others?

Separation of desired DNA from all others is done through the process of purification

using microfilter or gel and running the sample in a microconcentrator column by

centrifuge process. The process will trap the PCR products or the desired DNA, setting

aside the materials that are not needed in the collection tubes for discard which may

contain primers, nucleotides, and other undesired small compounds. The column

containing the PCR products will be then attached to the inverted tube and once again

centrifuged after adding buffers to loosen the DNA. The collection tube will then contain

mostly 1,500bp-long 16S rDNA with little contamination of long-stranded DNA (Black,

2012).
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4. How does an automatic DNA sequencer work?

A DNA sequencer is a scientific instrument used to automate the DNA

sequencing process. This scientific instrument runs a sample of DNA and determines the

order of the four bases: G (guanine), C (cytosine), A (adenine) and T (thymine). This is

then reported as a text string, called a read. This order of the four bases in a DNA

sequence that can be used to identify bacteria since each sequence of DNA is unique to a

particular species (Black, 2012).

5. Why is it possible to use a DNA sequence to identify bacteria?

Every bacterium has a unique DNA sequence. By studying the pattern of the sequence

information in each tube, the bacteria will be identified by matching the sequence of their

16SrDNA to the DNA sequence of the other previously identified bacteria with known

sequences in the database. The use of tools like Basic Local Alignment Search Tool

(BLAST) can facilitate the matching process. The sequence is unique to every species

making the identification of the bacteria possible by matching the pattern of the DNA

sequence. Perfect match DNA sequence would indicate the bacteria is identical to that in

the database while a sequence pattern with some differences may indicate a variation of

species or could be a new species which needs further assessment (Black, 2012).
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References

Black, J. (2012). Microbiology: Principles and explorations. (8th ed.) Hoboken, NY