Publication: Comparative analysis of DNA extraction methods in the recovery of human DNA from earphones
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Date
2026-02
Authors
Ying, Leeanne Yong Luo
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Abstract
Personal devices like earphones can be significant in forensic investigations as a source of DNA, especially when biological evidence is lacking. Although several studies have demonstrated successful DNA extraction and profiling from earphone samples, the most suitable DNA extraction method remains undescribed. DNA extraction is one of the most crucial steps in DNA analysis. Therefore, this study aimed to compare three different extraction methods, including silica-based commercial kit, Chelex-100 and phenol-chloroform method, to identify the most efficient method in recovering human DNA from earphones. A total of 66 samples were analysed, with 22 samples extracted using each method. DNA yield and purity were assessed using DeNovix DS-11, followed by PCR amplification using hGH autosomal primer and mitochondrial primer to assess DNA integrity. 3 samples from each method were further subjected to Real-time qPCR for quantification of amplifiable DNA. The findings demonstrated that phenol-chloroform yielded the highest and fluctuating DNA concentrations, as measured by DeNovix DS-11; however, the lowest yield when measured by qPCR, indicating contamination and overestimation by the spectrophotometer. The silica-based method yielded the lowest concentrations in spectrophotometry measurement, and moderate concentrations in the qPCR, but was the only method with successful autosomal amplification. The Chelex method displayed the second-highest concentration, but the highest concentration in qPCR. Chelex also demonstrated the greatest amplification success rate using mtDNA. These findings indicate that no single extraction method can excel in both quantity and quality parameters simultaneously. However, the Chelex method was found to recover DNA that performed better than the other methods, in terms of yield and amplifiability
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