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You’re working in a research lab, and your current task is to clone the gene that codes for tyrosinase from potatoes. You grind up some potato, extracts the DNA from it and digests the DNA with two different restriction enzymes (separately, not together): EcoRI and BamHI.  You then obtain the cloning vector, pUC19, and digest it with the same two enzymes.  You then run a gel which is shown here.  You notice that the cloning vector made nice, tight bands on the gel, but the potato DNA just looks like a smear with no distinct bands.  However, this is just what you expected.  Explain why there are so many bands. Which enzyme would be the better choice to use for cloning the potato DNA, EcoRI, or BamHI? Explain why?  Be specific

You’re working in a research lab, and your current task is to clone the gene that codes for tyrosinase from potatoes. You grind up some potato, extracts the DNA from it and digests the DNA with two different restriction enzymes (separately, not together): EcoRI and BamHI.  You then obtain the cloning vector, pUC19, and digest it with the same two enzymes.  You then run a gel which is shown here.  You notice that the cloning vector made nice, tight bands on the gel, but the potato DNA just looks like a smear with no distinct bands.  However, this is just what you expected.  Explain why there are so many bands. Which enzyme would be the better choice to use for cloning the potato DNA, EcoRI, or BamHI? Explain why?  Be specific

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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You’re working in a research lab, and your current task is to clone the gene that codes for tyrosinase from potatoes. You grind up some potato, extracts the DNA from it and digests the DNA with two different restriction enzymes (separately, not together): EcoRI and BamHI.  You then obtain the cloning vector, pUC19, and digest it with the same two enzymes.  You then run a gel which is shown here. 

    1. You notice that the cloning vector made nice, tight bands on the gel, but the potato DNA just looks like a smear with no distinct bands.  However, this is just what you expected.  Explain why there are so many bands.

    2. Which enzyme would be the better choice to use for cloning the potato DNA, EcoRI, or BamHI? Explain why?  Be specific.

The image displays a diagram of the pUC19 plasmid map, which is a circular DNA molecule commonly used in molecular cloning. Key Features of the pUC19 Plasmid: 1. **Ampicillin Resistance Gene (Amp^r)**: This provides resistance to the antibiotic ampicillin, allowing for the selection of bacteria that have successfully incorporated the plasmid. 2. **Polylinker (Multiple Cloning Site, MCS)**: Located between base pairs 396-454, the polylinker is a sequence containing multiple restriction enzyme sites. This allows for the insertion of foreign DNA at various sites, facilitating cloning. 3. **Replication Origin (ori)**: Not explicitly labeled on the diagram, but it is a necessary component for plasmid replication within bacterial cells. 4. **LacZ Gene**: Part of the plasmid that encodes for β-galactosidase, used for blue/white screening in cloning experiments. The plasmid is depicted as a circular structure, illustrating its closed-loop configuration, which is typical for plasmids. The diagram highlights the polylinker region in blue, emphasizing its significance in cloning applications.
Transcribed Image Text:The image displays a diagram of the pUC19 plasmid map, which is a circular DNA molecule commonly used in molecular cloning. Key Features of the pUC19 Plasmid: 1. **Ampicillin Resistance Gene (Amp^r)**: This provides resistance to the antibiotic ampicillin, allowing for the selection of bacteria that have successfully incorporated the plasmid. 2. **Polylinker (Multiple Cloning Site, MCS)**: Located between base pairs 396-454, the polylinker is a sequence containing multiple restriction enzyme sites. This allows for the insertion of foreign DNA at various sites, facilitating cloning. 3. **Replication Origin (ori)**: Not explicitly labeled on the diagram, but it is a necessary component for plasmid replication within bacterial cells. 4. **LacZ Gene**: Part of the plasmid that encodes for β-galactosidase, used for blue/white screening in cloning experiments. The plasmid is depicted as a circular structure, illustrating its closed-loop configuration, which is typical for plasmids. The diagram highlights the polylinker region in blue, emphasizing its significance in cloning applications.
**Gel Electrophoresis in DNA Fingerprinting** **Introduction** Gel electrophoresis is a key technique used in molecular biology to separate DNA, RNA, or proteins based on their size. In DNA fingerprinting, this method allows the visualization of DNA fragments. **Diagram Explanation** 1. **Structure** - The diagram illustrates a gel electrophoresis setup with DNA samples loaded into wells at the top. - A voltage is applied that moves DNA fragments towards the positive electrode due to the negatively charged backbone of DNA. 2. **Lane Components** - Far left: DNA ladder or marker with known fragment sizes, serving as a reference for estimating the size of other fragments. - Middle lanes: DNA samples that have been processed to reveal unique band patterns. - These bands represent DNA fragments of different lengths. 3. **Interpreting the Gel** - Bands closer to the wells are larger DNA fragments due to slower movement through the gel matrix. - Bands farther from the wells are smaller DNA fragments due to quicker movement. - Unique patterns in lanes are used to distinguish between different DNA samples, essential in DNA fingerprinting for applications such as criminal investigations, paternity testing, and genetic studies. **Conclusion** The gel's banding pattern provides a visual representation of the DNA's molecular size distribution. By comparing patterns, it is possible to determine genetic similarities or differences across samples, enabling identification and analysis in various research and practical applications.
Transcribed Image Text:**Gel Electrophoresis in DNA Fingerprinting** **Introduction** Gel electrophoresis is a key technique used in molecular biology to separate DNA, RNA, or proteins based on their size. In DNA fingerprinting, this method allows the visualization of DNA fragments. **Diagram Explanation** 1. **Structure** - The diagram illustrates a gel electrophoresis setup with DNA samples loaded into wells at the top. - A voltage is applied that moves DNA fragments towards the positive electrode due to the negatively charged backbone of DNA. 2. **Lane Components** - Far left: DNA ladder or marker with known fragment sizes, serving as a reference for estimating the size of other fragments. - Middle lanes: DNA samples that have been processed to reveal unique band patterns. - These bands represent DNA fragments of different lengths. 3. **Interpreting the Gel** - Bands closer to the wells are larger DNA fragments due to slower movement through the gel matrix. - Bands farther from the wells are smaller DNA fragments due to quicker movement. - Unique patterns in lanes are used to distinguish between different DNA samples, essential in DNA fingerprinting for applications such as criminal investigations, paternity testing, and genetic studies. **Conclusion** The gel's banding pattern provides a visual representation of the DNA's molecular size distribution. By comparing patterns, it is possible to determine genetic similarities or differences across samples, enabling identification and analysis in various research and practical applications.
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