Bacterial ID lab

1. As the medical technician in charge of this investigation, what are you trying to determine about the tissue sample provided to you?

I am trying to determine the sequence and the identity of the tissue sample

2. How did you prepare the DNA to be used in this investigation?

Extract the bacterial colony, use a small pipette and transfer it into a micro centrifuge tube, then put it into a heating bath and put it into a centrifuge after that transfer it into a PCR tube.

3. Describe how PCR is used to make copies of DNA sequences. Use the animation and notebook entries in the PCR Amplification step to guide your answer. Note that you may replay the animation as needed.

The heat separates the DNA then it cools it and the DNA makes a copy then it repeats over and over again.

4. Summarize the technique used to purify the PCR product.

Use a buffer solution put it into the column then add PCR, the Other PCR tubes are cooling while you load the column  into the centrifuge for 15 minutes then invert the column into a new column and add more buffer solution into it get rid of the 1st column and the 2nd column will be put into a centrifuge wait 2min then get rid of the column and now the PCR product has been purified.

5. What is produced during the sequencing prep PCR run? Use the animation and notebook as needed in thinking through your answer. 
The DNA pieces in each tube start in the same primer but end with a different nucleotide showing the construction of the DNA.

6. Describe how the automatic sequencer determines the sequences of the PCR products.
Performs gel electrophoresis on the DNA in each tube separating molecules

based on their size

7. What does BLAST stand for? BLAST stands for Basic Local Alignment Search Tool

8. What conclusions did you make using the results of the BLAST search?  Did these conclusions support a clinical diagnosis for the patient (what disease did they have)?
I got a disease called Bartonella Henselae and it did support a clinical diagnosis for the patient.

Bartonella Henselae: Various species of Bartonella that are pathogenic to humans are transmitted via a vector, or directly from an animal reservoir. For example, B. bacilliformis via sandflies causes Oroya fever; B. quintana via body lice causes trench fever; and B. henselae via cats causes cat scratch disease (CSD). CSD typically manifests as swellings of the lymph glands, possibly with skin lesions at the site of inoculation and possibly accompanied by fever, fatigue, and other symptoms. Immunocompromised patients may be particularly susceptible and can develop a different disease, bacillary angiomatosis, as a result of infection by B. henselae or B. quintana.