Pre-lab (Week 7)

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Skyline College *

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253L

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Biology

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Dec 6, 2023

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docx

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2

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Pre-lab questions for Biol 253L, week 7. The answers are found in this week’s reading. Answer in the space below and continue on back if necessary. 1. Suppose that when you come into lab this week and expose the bacterial colonies on your LB Agar + Ampicillin plate to UV light, you do not observe any green fluorescence. What is the most logical conclusion? The most logical conclusion would be that the plasmid does not contain GFP if there is no green fluorescence present. 2. Do you expect your LB Agar + Ampicillin plate to have more, fewer, or the same number of bacterial colonies compared to your LB Agar plate? Explain your answer. LB Agar + Ampicillin would have fewer bacterial colonies due to ampicillin being able to kill/resist infections compared to LB Agar plate. 3. At neutral pH, is DNA negatively charged, positively charged, or uncharged? If DNA is charged, state which chemical group in the DNA makes it charged. At neutral pH, DNA has a negative charge because phosphate groups are present. 4. Suppose you have DNA fragments that are 1000 bp and DNA fragments that are 500 bp. If you perform gel electrophoresis with these DNA fragments, which ones would migrate faster through the gel? Explain. 500 bp DNA fragments would migrate faster through the gel than the 1000 bp DNA fragments since the attractive force from the electric charge and the force of the gel is affected by the size of the DNA fragment. 5. Why is ethidium bromide used in gel electrophoresis? Ethidium bromide is a fluorescent dye used in gel electrophoresis to make it visible. 6. What is the purpose of using a DNA ladder in gel electrophoresis? The purpose of using a DNA ladder in gel electrophoresis is to determine the sizes of DNA molecules in the other lanes by comparing the positions of the bands to those in the reference lane. 7. True or false: Two different DNA molecules that both have a molecular weight of 2000 bp should always migrate at the same speed in gel electrophoresis. Explain, giving an example to justify your answer. For example, if two different DNA molecules that have a weight of 400 bp it would migrate at the same speed. If one was 400 bp and the other was 300 bp, one would be slower than the other. 8. What will you add to your restriction digests just before loading them on the gel?
4 l of loading dye will be added to each of your three restriction digest tubes as well as the uncut plasmid control tubes. 9. When collecting your P1 and P2 Fast Plant leaves for freezing, should the leaves come from plants in the same pot or in different pots? The leaves should come from different pots.
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