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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 Isolation01:34

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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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Simultaneous DNA-RNA Extraction from Coastal Sediments and Quantification of 16S rRNA Genes and Transcripts by Real-time PCR
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Sticky problems: extraction of nucleic acids from molluscs.

Coen M Adema1

  • 1Center for Evolutionary and Theoretical Immunology, Department of Biology, University of New Mexico, Albuquerque, NM 87112, USA.

Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences
|April 5, 2021
PubMed
Summary

Extracting DNA and RNA from molluscs is challenging due to their unique biology. Hexadecyltrimethylammoniumbromide (CTAB) methods improve DNA recovery, while RNA analysis requires specialized techniques for accurate molecular malacology studies.

Keywords:
AT richCTABhidden breakmolluscamucopolysaccharidespigment

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Area of Science:

  • Molecular biology
  • Malacology
  • Genomics

Background:

  • Molluscan taxonomy and biology benefit from molecular and omics techniques.
  • Diverse molluscan biological properties, including biomineralization and mucus, complicate nucleic acid recovery.
  • Polyphenolic proteins and mucopolysaccharides interfere with DNA extraction and molecular procedures.

Purpose of the Study:

  • To address challenges in DNA and RNA extraction from molluscs for molecular studies.
  • To highlight methods facilitating molecular-level investigations in Mollusca.
  • To inform the expansion of molecular malacology, including work with museum specimens and next-generation sequencing.

Main Methods:

  • Utilizing hexadecyltrimethylammoniumbromide (CTAB) detergent for DNA extraction to remove contaminants.
  • Employing gel electrophoresis or fluorometry for accurate DNA quantification due to pigment interference.
  • Adapting RNA extraction protocols to account for the 'hidden break' in 28S rRNA and high-AT content.

Main Results:

  • CTAB-based DNA extraction methods effectively facilitate molecular studies in molluscs.
  • Spectrophotometry is unreliable for DNA quantification; gel electrophoresis or fluorometry are necessary.
  • Standard RNA quality control methods (18S:28S rRNA ratio) are not directly applicable to molluscs.

Conclusions:

  • Understanding molluscan-specific challenges in nucleic acid extraction is crucial for advancing molecular malacology.
  • Modified methods enable successful molecular analyses, including next-generation sequencing, from diverse molluscan samples.
  • Continued development of extraction techniques is vital for exploring the genomics of this neglected phylum.