Module 5 BACTERIAL IDENTIFICATION VIRTUAL LAB

BACTERIAL IDENTIFICATION VIRTUAL LAB worksheet.pdf

BACTERIAL IDENTIFICATION VIRTUAL LAB

https://www.biointeractive.org/classroom-resources/bacterial-identification-virtual-lab (Links to an external site.)

Introduction

1. What is the overall purpose of this virtual lab?
2. What are the four basic steps involved in this bacterial identification lab?
3. What are the differences between a prokaryotic and a eukaryotic ribosome?
4. What is aSvedberg unit?
5. What is 16s rRNA and why is it useful in classifying bacteria?

 PART 1: SAMPLE PREPARATION

1. Extracting DNA involves which initial step?
2. heating the sample in a water bath at 100°C
3. Is the DNA in the supernatant (the liquid),or the pellet, after centrifugation?

PART 2: PCR AMPLIFICATION

1. List each step of a PCR cycle, the temperature, and the duration (time).
2. What are the components of the PCR Master Mix solution?
3. What are primers?
4. What are the negative and positive controls used in this experiment?

PCR PURIFICATION

1. Approximately how long is the 16s rDNA (bp)?
2. If you were to run a gel, it would have three lanes. What would each lane contain, and what would you see in each lane after running the gel?
3. In order to purify the PCR product, you use a microconcentrator column.What should the final collection tube contain?

PART 4: SEQUENCING PREPARATION

1. List the steps in cycle sequencing
2. Describe the “sequencing brew” to which you added your purified PCR product.

PART 5: DNA SEQUENCING

1. What is the final PCR product, the material contained in your 12 tubes?
2. What is the purpose of the laser beam in determining a DNA sequence?

PART 6: DNA SEQUENCE ANALYSIS

1. What is the ultimate goal of the sequence matching analysis?
2. What is “homology”?
3. What is BLAST and how is it used?
4. After selecting and copying the sequence data inside the gray box and performing the BLAST search, what was the scientific name of the bacterium you sequenced?
5. Write a brief description of this bacterium
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