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Acetone preservation: a practical technique for molecular analysis.
1National Institute of Bioscience and Human Technology, Agency of Industrial Science and Technology, Tsukuba, Ibaraki, Japan. fukatsu@nibh.go.jp
Molecular Ecology
|January 5, 2000
Summary
Acetone effectively preserves insect and symbiont DNA, RNA, and proteins for long-term molecular analysis, outperforming ethanol, especially with water contamination. This method is ideal for field collection and archival preservation.
Area of Science:
- Molecular Biology
- Entomology
- Microbiology
Background:
- Field collection and preservation of biological samples for molecular analysis present challenges.
- Existing methods using ethanol may be suboptimal for preserving DNA, RNA, and proteins, particularly in the presence of water.
Purpose of the Study:
- To evaluate the efficacy of various organic solvents, including acetone and ethanol, for the preservation of insect and endosymbiont DNA, RNA, and proteins.
- To identify a reliable and convenient method for field collection and archival storage of specimens for molecular analyses.
Main Methods:
- Insects (pea aphid Acyrthosiphon pisum) and their symbionts were stored in various organic solvents (acetone, ethanol, 2-propanol, diethyl ether, ethyl acetate, methanol, chloroform).
- DNA, RNA, and protein integrity were assessed using polymerase chain reaction (PCR), gel electrophoresis (SDS-PAGE), immunoblotting, immunohistochemistry, and in situ hybridization after varying storage periods (6 months to several years).
- The impact of water contamination on solvent efficacy was specifically tested for acetone and ethanol.
Main Results:
- Acetone, ethanol, 2-propanol, diethyl ether, and ethyl acetate effectively preserved DNA and rRNA after 6 months.
- Acetone demonstrated superior performance compared to ethanol in preserving DNA, particularly when water was present.
- Long-term storage (several years) in acetone maintained the integrity of insect and endosymbiont DNA, RNA, and proteins, enabling successful molecular analyses.
Conclusions:
- Acetone is a highly effective and recommended solvent for the long-term preservation of insect and endosymbiont DNA, RNA, and proteins for molecular analysis.
- Acetone offers advantages over ethanol, especially in field conditions where water contamination is likely.
- This method facilitates reliable field collection and archival preservation of diverse insect-associated microbial communities.