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Effects of different preservation methods on inter simple sequence repeat (ISSR) and random amplified polymorphic DNA
Xiaolong Wang1, Lin Li1, Jiaxin Zhao1
1National & Local United Engineering Laboratory for Chinese Herbal Medicine Breeding and Cultivation, Jilin University, Changchun, Jilin Province 130012, People's Republic of China; School of Life Sciences, Jilin University, Changchun, Jilin Province 130012, People's Republic of China.
Comptes Rendus Biologies
|April 5, 2017
Summary
Sample preservation methods significantly impact DNA analysis for molecular markers. Storing plant samples in liquid nitrogen or silica gel is effective for inter simple sequence repeat (ISSR) and random amplified polymorphic DNA (RAPD) analyses, with minimal time effects.
Area of Science:
- Plant molecular biology
- Genetics
- Biobanking
Background:
- Accurate DNA preservation is crucial for molecular marker analysis.
- Various methods exist for preserving botanical specimens, but their impact on DNA quality and downstream analyses is not fully understood.
- Inter simple sequence repeat (ISSR) and random amplified polymorphic DNA (RAPD) are widely used molecular markers.
Purpose of the Study:
- To assess the influence of different preservation techniques on DNA quality and subsequent ISSR and RAPD analyses.
- To compare the effectiveness of -20°C ice chest, liquid nitrogen, and silica gel drying for preserving diverse plant types.
- To determine the impact of short-term (three weeks) versus long-term (three years) preservation on molecular marker data.
Main Methods:
- Statistical analysis of DNA band numbers, genetic indices, and cluster analysis.
- Evaluation of DNA yield and quality from preserved samples.
- Application of ISSR and RAPD marker techniques on DNA extracted from preserved specimens.
Main Results:
- All tested preservation methods yielded sufficient genomic DNA for ISSR and RAPD analyses.
- Preservation method significantly affected the outcomes of ISSR and RAPD analyses.
- Preservation duration (three weeks vs. three years) had a minimal impact on the results.
- Effectiveness varied among preservation methods depending on plant type (broad-leaved, needle-leaved, succulent).
Conclusions:
- Preservation method is a critical factor influencing molecular marker analysis outcomes.
- Liquid nitrogen and silica gel drying are generally superior to -20°C ice chest storage for DNA-based molecular marker studies.
- Researchers should select preservation methods based on their specific plant material and analytical goals to ensure reliable genetic data.