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Complete the following problems. Restriction enzymes (REs), which cut D NA at specific sequences, are classic tools in molecular biology. Because of their specificity in cutting DNA, REs can be used to "map" DNA sequences by analyzing the fragments generated upon restriction digest, as in the example shown in Figure 1. Your task is to study the circular plasmid, PMBBS, through restriction digests. You subjected the PMBBS plasmid to complete digestion by different combinations of three REs (ECORI, BamHI, and Xhol), and analyzed the results on an agarose gel, shown below. Using the data you can glean from this gel, answer the questions that follow. EcoRI BamHI Xhol DNA size ladder 3000 bp 2500 bp 2000 bp 1500 bp 1200 bp 1000 bp 900 bp 800 bp 700 bp 600 bp 500 bp 400 bp 300 bp 200 bp 100 bp "vC 100bp Plus DNA ladder from Vivantis Techvnologies "The same total amount of DNA was loaded in each lane. 1. What is the total size of the PMBBS plasmid in bp? Answer bp 2. How many cut sites on the pMBBS plasmid does each RE have? EcoRI:, BamHI:, Xhol: 3. If the pMBBS plasmid were digested with all three REs together, how many fragments would be observed, and what would be the sizes (in bp) of these fragments? Number of fragments: Sizes:

Complete the following problems. Restriction enzymes (REs), which cut D NA at specific sequences, are classic tools in molecular biology. Because of their specificity in cutting DNA, REs can be used to "map" DNA sequences by analyzing the fragments generated upon restriction digest, as in the example shown in Figure 1. Your task is to study the circular plasmid, PMBBS, through restriction digests. You subjected the PMBBS plasmid to complete digestion by different combinations of three REs (ECORI, BamHI, and Xhol), and analyzed the results on an agarose gel, shown below. Using the data you can glean from this gel, answer the questions that follow. EcoRI BamHI Xhol DNA size ladder 3000 bp 2500 bp 2000 bp 1500 bp 1200 bp 1000 bp 900 bp 800 bp 700 bp 600 bp 500 bp 400 bp 300 bp 200 bp 100 bp "vC 100bp Plus DNA ladder from Vivantis Techvnologies "The same total amount of DNA was loaded in each lane. 1. What is the total size of the PMBBS plasmid in bp? Answer bp 2. How many cut sites on the pMBBS plasmid does each RE have? EcoRI:, BamHI:, Xhol: 3. If the pMBBS plasmid were digested with all three REs together, how many fragments would be observed, and what would be the sizes (in bp) of these fragments? Number of fragments: Sizes:

Biochemistry
Biochemistry
6th Edition
ISBN:9781305577206
Author:Reginald H. Garrett, Charles M. Grisham
Publisher:Reginald H. Garrett, Charles M. Grisham
Chapter28: Dna Metabolism: Replication, Recombination, And Repair
Section: Chapter Questions
Problem 1P: Semiconservative or Conservative DNA Replication If 15N-Iabeled E. coli DNA has a density of 1.724...
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l MTN ? 13:30 © 0 8% O Google Drive - online backup Open in the Drive app ОPEN Complete the following problems. Restriction enzymes (RES), which cut D NA at specific sequences, are classic tools in molecular biology. Because of their specificity in cutting DNA, REs can be used to "map" DNA sequences by analyzing the fragments generated upon restriction digest, as in the example shown in Figure 1. Your task is to study the circular plasmid, pMBBS, through restriction digests. You subjected the PMBBS plasmid to complete digestion by different combinations of three RES (ECORI, BamHI, and Xhol), and analyzed the results on an agarose gel, shown below. Using the data you can glean from this gel, answer the questions that follow. EcoRI BamHI Xhol DNA size ladder 3000 bp 2500 bp 2000 bp 1500 bp 1200 bp 1000 bp 900 bp 800 bo 700 bp 600 bp 500 bp 400 bp 300 bp - 200 bp 100 bo vC 100bp Plus DNA ladder from Vivantis Technologies "The same total amount of DNA was loaded in each lane. 1. What is the total size of the PMBBS plasmid in bp? Answer: bp 2. How many cut sites on the PMBBS plasmid does each RE have? BamHI: EcoRI: Xhol: 3. If the pMBBS plasmid were digested with all three REs together, how many fragments would be observed, and what would be the sizes (in bp) of these fragments? Number of fragments: Sizes:
Transcribed Image Text:l MTN ? 13:30 © 0 8% O Google Drive - online backup Open in the Drive app ОPEN Complete the following problems. Restriction enzymes (RES), which cut D NA at specific sequences, are classic tools in molecular biology. Because of their specificity in cutting DNA, REs can be used to "map" DNA sequences by analyzing the fragments generated upon restriction digest, as in the example shown in Figure 1. Your task is to study the circular plasmid, pMBBS, through restriction digests. You subjected the PMBBS plasmid to complete digestion by different combinations of three RES (ECORI, BamHI, and Xhol), and analyzed the results on an agarose gel, shown below. Using the data you can glean from this gel, answer the questions that follow. EcoRI BamHI Xhol DNA size ladder 3000 bp 2500 bp 2000 bp 1500 bp 1200 bp 1000 bp 900 bp 800 bo 700 bp 600 bp 500 bp 400 bp 300 bp - 200 bp 100 bo vC 100bp Plus DNA ladder from Vivantis Technologies "The same total amount of DNA was loaded in each lane. 1. What is the total size of the PMBBS plasmid in bp? Answer: bp 2. How many cut sites on the PMBBS plasmid does each RE have? BamHI: EcoRI: Xhol: 3. If the pMBBS plasmid were digested with all three REs together, how many fragments would be observed, and what would be the sizes (in bp) of these fragments? Number of fragments: Sizes:
l MTN 13:30 O 8% Google Drive - online backup Open in the Drive app OPEN 4. Draw the actual map of the PMBBS plasmid, following the style of the sample map shown in Figure 1. Make sure you show the following information: (1) relative locations of the RE sites; (2) size in base pairs of the segments of DNA between sites. (The map need not be drawn to scale.) Answer: (provide a drawing)
Transcribed Image Text:l MTN 13:30 O 8% Google Drive - online backup Open in the Drive app OPEN 4. Draw the actual map of the PMBBS plasmid, following the style of the sample map shown in Figure 1. Make sure you show the following information: (1) relative locations of the RE sites; (2) size in base pairs of the segments of DNA between sites. (The map need not be drawn to scale.) Answer: (provide a drawing)
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