5 Questions
Which method is suitable for clearing huge amounts of debris but takes more time?
Centrifugation
Which method can be used in automated protocols but gets overwhelmed with biomass?
Bead-based clearing
Which molecular separation method involves Agarose Gel Electrophoresis?
Centrifugation
Which matrices can be used for binding to genomic DNA for purification?
Silica, cellulose, and ion exchange matrices
Which method selectively binds to DNA, leaving out RNA and proteins?
Purification Binding Matrix
Study Notes
DNA Extraction Methods
- Physical method: involves creating a lysate from tough tissues through freezing of cells, grinding with mortar and pestle under liquid nitrogen, and sonication using metallic or ceramic beads.
- Chemical method: applies to easy-to-lyse materials like tissue-culture cells, and involves using various agents for disruption of cell membranes and proteins, such as detergents like SDS and chaotropes like alkaline solutions and guanidine salts.
- Enzymatic method: used for disruption of plant materials, tissues, bacteria, and yeast, and involves enzymes that disrupt tissues and tough cell walls, including lysozyme, liticase, zymolyase, collagenase, lipase, and proteinase K.
Lysate Clearing
- Unwanted materials in the lysate, such as lipids, proteins, and saccharides, can clog with membranes or interfere with downstream applications during the purification reaction.
- Clearing is established through filtration, centrifugation, or bead-based methods to remove these unwanted materials from the lysate.
Test your knowledge of DNA extraction methods with this quiz. Explore physical, chemical, enzymatic methods, and combinations of the three. Learn about creating lysate from tough tissues using freezing, grinding, and sonication, as well as chemical methods for easy-to-lyse materials.
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