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Nucleic Acids02:43

Nucleic Acids

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Nucleic acids are the most important macromolecules for the continuity of life. They carry the cell's genetic blueprint and carry instructions for its functioning.
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The two main types of nucleic acids are deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). DNA is the genetic material in all living organisms, ranging from single-celled bacteria to multicellular mammals. It is in the nucleus of eukaryotes and in the organelles, chloroplasts, and mitochondria. In prokaryotes,...
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Nucleic acids are the most important macromolecules for the continuity of life. They carry the cell's genetic blueprint and carry instructions for its functioning.
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Nucleic Acid Structure01:25

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The pentose sugar in DNA is deoxyribose, while in RNA the pentose sugar is ribose. The difference between the sugars is the presence of the hydroxyl group on the ribose's second carbon and a hydrogen on the deoxyribose's second carbon. The phosphate residue attaches to the hydroxyl group of the 5′ carbon of one sugar and the hydroxyl group of the 3′ carbon of the sugar of the next nucleotide, which forms  a 5′ to 3′ phosphodiester linkage.
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Nucleic Acids and Nucleotides01:20

Nucleic Acids and Nucleotides

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Nucleic acids are the most important macromolecules for the continuity of life. They carry the cell's genetic blueprint and have instructions for its functioning. The two main types of nucleic acids are deoxyribonucleic acid (DNA) and ribonucleic acid (RNA).
Deoxyribonucleic Acid (DNA)
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Biosynthesis of Nucleic Acids01:28

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Nucleic acid biosynthesis is a fundamental biochemical process that produces the purine and pyrimidine nucleotides essential for DNA and RNA synthesis. This pathway maintains a balanced nucleotide pool, preventing imbalances that could jeopardize genetic integrity and cellular function. Given the crucial role of nucleotides, their synthesis is tightly regulated to ensure proper cellular homeostasis.Purine BiosynthesisThe biosynthesis of purine nucleotides begins with ribose-5-phosphate, a...
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Kinetic Screening of Nuclease Activity using Nucleic Acid Probes
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Nucleic Acid Extraction from Human Biological Samples.

Sureni V Mullegama1, Michael O Alberti1, Cora Au1

  • 1Department of Pathology and Laboratory Medicine, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|December 13, 2018
PubMed
Summary
This summary is machine-generated.

High-quality nucleic acid isolation is crucial for biomedical research. This guide details DNA and RNA extraction from various tissues, including saliva and FFPE samples, with troubleshooting tips.

Keywords:
AmniocyteBloodBone marrow aspirateBuccal cellsCultured cellsDNA extractionFormalin-fixedFresh/frozen tissueNucleic acidsParaffin-embedded (FFPE) tissueRNASaliva

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Area of Science:

  • Molecular Biology
  • Biomedical Research

Background:

  • Nucleic acid isolation is a foundational step in molecular biology techniques.
  • Standardized reagents and commercial kits are widely available for DNA and RNA extraction.

Purpose of the Study:

  • To provide a comprehensive overview of nucleic acid extraction protocols.
  • To address challenges and offer solutions for various sample types, including challenging ones like saliva and FFPE tissues.
  • To discuss essential steps for successful nucleic acid isolation and quantitation.

Main Methods:

  • Detailed protocols for DNA and RNA extraction from diverse biological samples.
  • Discussion of critical steps: tissue disruption, nucleoprotein denaturation, nuclease inactivation, and purification.
  • Consideration of specific sample types such as saliva and formalin-fixed, paraffin-embedded (FFPE) tissues.
  • Exploration of common challenges and pitfalls in nucleic acid isolation.
  • Overview of nucleic acid quantitation techniques.

Main Results:

  • Successful nucleic acid extraction relies on effective tissue disruption, denaturation, nuclease inactivation, and purification.
  • Protocols can be adapted based on the nucleic acid of interest and the biological sample source.
  • Specific considerations are necessary for challenging samples like saliva and FFPE tissues.
  • Common pitfalls can be identified and mitigated through careful protocol adherence.

Conclusions:

  • Effective nucleic acid isolation is paramount for reliable downstream molecular experiments.
  • This chapter provides essential guidance for researchers working with diverse sample types.
  • Understanding and addressing potential pitfalls ensures high-quality nucleic acid yield and purity.