iscovered that a species of bacteria can break down Styrofoam™ (polystyrene) products due to an enzyme it produces, polystyrenase. You wish to study the gene that codes for this enzyme. Task 1: DNA Extraction To begin work on the bacterium, you begin by extracting its genomic DNA (gDNA). What is the purpose of the following procedures? Answer briefly but completely. Using sodium dodecyl sulfate, a detergent Answer: __________________________________
You discovered that a species of bacteria can break down Styrofoam™ (polystyrene) products due to an enzyme it produces, polystyrenase. You wish to study the gene that codes for this enzyme.
Task 1: DNA Extraction
To begin work on the bacterium, you begin by extracting its genomic DNA (gDNA). What is the purpose of the following procedures? Answer briefly but completely.
- Using sodium dodecyl sulfate, a detergent
Answer: __________________________________
- Adding RNase A and Proteinase K during extraction
Answer: __________________________________
- Adding ethanol before recovering the DNA extract
Answer: __________________________________
Task 2: Polymerase Chain Reaction
After purifying the gDNA extract, you want to isolate and amplify the polystyrenase gene. You perform PCR using the appropriate gene-targeted primers. What is the purpose of the following PCR components? Answer briefly but completely.
- DNA polymerase isolated from Thermus aquaticus
Answer: __________________________________
- Deoxynucleotide triphosphates (dNTPs)
Answer: __________________________________
- Forward and reverse primers
Answer: __________________________________
Task 3: Agarose Gel Electrophoresis
Your amplicon from PCR was subjected to AGE for analysis. You are shown the image of the gel loaded with the following lanes: (A) negative control, (B) size ladder (1, 2, 3, 4, 6, and 10 kb), (C) gDNA extract, (D) PCR amplicon. However, due to mishaps while loading the gel with the samples, you are not sure which lane is which. You are shown a diagram of the obtained gel below.
- Label each lane of the gel. Write only the corresponding letters in the wells above.
- Above each band in the size ladder, write its size (in kb).
- Approximate the size (in kb) of the polystyrenase gene. Write your answer above the band corresponding to the gene.
Bonus: If you wish to identify the bacterial species in this scenario, what gene is most commonly and routinely sequenced?
Answer: __________________________________
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