Class 12 · Biotechnology: Principles and Processes

DNA Isolation and Purification Methods — NEET Biology

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Recall that nucleic acid is the genetic material of all organisms without exception. In majority of organisms this is deoxyribonucleic acid or DNA. In order to cut the DNA with restriction enzymes, it needs to be in pure form, free from other macro-molecules. Since the DNA is enclosed within the membranes, we have to break the cell open to release DNA along with other macromolecules such as RNA, proteins, polysaccharides and also lipids. This can be achieved by treating the bacterial cells/plant or animal tissue with enzymes such as lysozyme (bacteria), cellulase (plant cells), chitinase (fungus). You know that genes are located on long molecules of DNA intertwined with proteins such as histones. The RNA can be removed by treatment with ribonuclease whereas proteins can be removed by treatment with protease. Other molecules can be removed by appropriate treatments and purified DNA ultimately precipitates out after the addition of chilled ethanol. This can be seen as collection of fine threads in the suspension.

🖼️Related NCERT figure: Two photographs showing DNA isolation process - top image shows DNA in a container, bottom image shows DNA being spooled out (Figure 9.5 DNA that separates out can be removed by spooling)
NCERT Biology · Class 12 · Chapter 9 · Paragraph 44
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Arrange the following steps of isolation and preparation of genetic material (DNA) in correct sequence: 1. Removal of RNA, proteins and other molecules by ribonuclease, protease and suitable treatments 2. Breaking open the cell wall using enzymes like lysozyme, cellulase or chitinase to release DNA 3. Precipitation of purified DNA as fine threads after addition of chilled ethanol 4. Recognition that nucleic acid (DNA) is the universal genetic material of all organisms 5. Releasing DNA along with RNA, proteins, polysaccharides and lipids from the disrupted cells

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