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Related Concept Videos

DNA Isolation01:24

DNA Isolation

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DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
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DNA from cells is required for many biotechnology and research applications, such as molecular cloning. To remove and purify DNA from cells, researchers use various methods of DNA extraction. While the specifics of different protocols may vary, some general concepts underlie the process of DNA extraction.
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DNA Agarose Gel Electrophoresis02:35

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Agarose gel electrophoresis is a laboratory technique commonly used to separate DNA fragments by size. However, it can also be used to isolate and purify DNA fragments using a gel extraction protocol.
Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA.
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Techniques for Isolation of Pure Cultures01:24

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Microorganisms are routinely cultured in the laboratory using various techniques to isolate, grow, and quantify them for further study. These methods rely on inoculating microorganisms into a suitable growth medium under aseptic conditions to prevent contamination. Depending on the objective, inoculation can involve direct transfer or the use of diluted bacterial suspensions as the inoculum.Streak-Plate Method for IsolationThe streak-plate method is a common technique for obtaining pure...
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Differential staining is an essential microbiological technique that exploits variations in cell wall structures to classify and identify microorganisms. It facilitates the distinction of bacteria, aiding in diagnostic and research applications. Two of the most widely used differential staining methods are Gram staining and acid-fast staining, both of which rely on the chemical and structural differences in bacterial cell walls.Gram Staining TechniqueGram staining differentiates bacteria by...
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Related Experiment Video

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Isolation, Propagation, and Identification of Bacterial Species with Hydrocarbon Metabolizing Properties from Aquatic Habitats
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Isolating DNA from Gram-Negative Bacteria.

Michael R Green, Joseph Sambrook

    Cold Spring Harbor Protocols
    |January 5, 2017
    PubMed
    Summary

    This protocol details bacterial DNA isolation using cell lysis with sodium dodecyl sulfate and proteinase K. The high-molecular-weight DNA is then purified for downstream applications.

    Area of Science:

    • Molecular Biology
    • Genomics
    • Biochemistry

    Background:

    • Bacterial DNA isolation is crucial for genetic analysis and molecular biology techniques.
    • Efficient extraction of high-quality bacterial DNA is essential for various research applications.

    Purpose of the Study:

    • To describe a reliable protocol for isolating bacterial DNA.
    • To provide a method for obtaining high-molecular-weight DNA from bacterial cells.

    Main Methods:

    • Bacterial cell lysis using sodium dodecyl sulfate and proteinase K.
    • Shearing of high-molecular-weight DNA to reduce viscosity.
    • Extraction with phenol:chloroform and precipitation with isopropanol.

    Main Results:

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  • Successful isolation of bacterial DNA.
  • Obtained DNA fragment lengths ranging from 30 to 80 kilobases (kb).

    • Conclusions:

    • The described protocol effectively yields high-molecular-weight bacterial DNA.
    • This method is suitable for applications requiring purified bacterial DNA.