Figure 2. This image represents DNA from wheat germ that was digested with micrococcal nuclease for 15 minutes and the resulting fragments were separated by agarose electrophoresis on a 1.5% gel. The amount of micrococcal nuclease added to the samples varied. Nucleosome Structure Data Analysis Lane: 1 2 3 1 Nucleosome+Linker ~165 bp Linker DNA ~50 bp 4. Referring to Figure 2: Was Lane 1 loaded with more or less sample as compared to lanes 2 and 3? Explain why. 5. Referring to Figure 2: As stated in the figure, each sample of DNA was digested for the same amount of time (15 minutes), but differing amounts of nuclease. Knowing this, which lane was digested with the largest amount of nuclease? Explain why. 6. Referring to Figure 2: How could you better visualize the shorter DNA fragments in land 2? What about the longer fragments in lane 3? 7. In a paragraph define nucleosome and what 5 you have learned about. 8. Explain how the charges on DNA and ethidium bromide effect their movement through the agarose gel during electrophoresis. Why is ethidium bromide used in electrophoresis?
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Referring to Figure 2: Was Lane loaded with more or less sample as compared to lanes and 37? Explain why. Since the figure is not provided, we cannot determine the specific lane. However, to compare the amount of sample loaded in different lanes, you can look at Show more…
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Figure 1. Questions 1-4. DNA from wheat germ was digested with micrococcal nuclease and the resulting fragments were separated by agarose electrophoresis on a 1.5% gel. Lanes 2, 3, 4 and 5 contain samples of DNA that were digested with 20 uL of nuclease. The length of the incubation time of the enzyme with the samples varied. Lane 1 contains a 100 bp ladder. 1. Please explain the dark smear in the upper portion of lane three. 2. Lane 3 appears to have a dark band in the well at the top of the gel, how do you explain this? 3. On the gel, label a band that contains DNA fragments that were wrapped around 2 nucleosomes plus the linker DNA [units should be in base pairs]. Approximately how many base pairs do you assume this to be, why? 4. This gel seems to have some poor resolution of gel bands in both lanes 2 and 3. Assuming that you have to run the samples together in a new gel, what could you do to optimize the resolution of the DNA fragments in lane 2 and lane 3?
Adi S.
The following figure shows your DNA agarose gel electrophoresis result (left panel) and the map of the recombinant plasmid called pcDNA3-SENP2 (right panel). The pcDNA3-SENP2 DNA was incubated at 37 °C in the absence (lane 2, left panel) or the presence (lane 3, left panel) of the restriction enzymes EcoRI and XbaI, followed by DNA gel electrophoresis. Based on the following figure, please estimate the size of the inserted SENP2 gene fragment released by the digestion using EcoRI and XbaI. As shown in the right panel, the size of the empty pcDNA3 vector (without insert DNA) is 5.4 kb. What is the size (kb) of the inserted SENP2 gene fragment? 1.8 (kb)
Madhur L.
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