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DNA Extraction and Comparison between Old and Fresh Necrophilic Fly Samples.

Bo Wang¹, Yu-Zhao Xu², Yun-Chao Zhou²

¹Hainan Province Tropical Forensic Engineering Research Center, Department of Forensic Medicine, Hainan Medical University, Hainan Provincial Academician Workstation (tropical forensic medicine); Hainan Vocational College of Political Science and Law, Hainan Gongping Forensic Center.

Journal of Visualized Experiments : Jove

|May 20, 2024

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DNA can be successfully extracted from necrophilous flies using a blood DNA extraction kit. Alcohol storage effectively preserves fly DNA longer than dry storage, maintaining higher DNA concentrations in older samples.

Area of Science:

Forensic entomology
Molecular biology
Genetics

Background:

Necrophilous flies, such as Chrysomya megacephala, are crucial in forensic investigations.
Preservation methods for insect samples can impact DNA quality and yield.
Efficient DNA extraction is vital for accurate genetic analysis.

Purpose of the Study:

To evaluate the efficacy of a commercial blood DNA extraction kit for extracting DNA from Chrysomya megacephala.
To assess the impact of different storage conditions (alcohol vs. dry storage) on DNA preservation over time.
To determine the optimal method for preserving necrophilous fly specimens for DNA analysis.

Main Methods:

DNA extraction from thorax tissues of fresh and aged Chrysomya megacephala samples using a blood DNA extraction kit.

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Assessment of DNA purity and concentration via microplate reader and fluorometer.

Polymerase Chain Reaction (PCR) amplification and electrophoresis using fly-specific primers targeting the mitochondrial CO I gene.

Main Results:

All tested samples yielded DNA with purity ratios greater than 2.0.
DNA concentration followed the order: fresh samples > alcohol-preserved samples > dry-stored samples.
Successful PCR amplification and electrophoresis were achieved for all samples, indicating viable DNA.

Conclusions:

Commercial blood DNA extraction kits are effective for obtaining DNA from necrophilous flies.
Alcohol preservation is a suitable long-term storage method for necrophilous flies, maintaining DNA integrity better than dry storage.
Freshly collected specimens yield higher DNA concentrations compared to aged specimens.