YFiler Platinum Casework kit performance was evaluated in the presence of PCR inhibitors such as hematin, humic acid, and tannic acid9 (link),10 (link). Stock solutions of each inhibitor compound were prepared as follows: hematin (Sigma, St. Louis, MO) was dissolved in 0.1 N NaOH; humic acid (Alfa Aesar) and tannic acid (Fisher Scientific) were dissolved in molecular biology grade distilled water (Ambion). Test samples were prepared with a constant input of 1 ng of 007 DNA per reaction and increasing concentrations of each inhibitor: 200 and 400 µM hematin, 100 and 200 ng/µL of humic acid, or 100, and 300 ng/µL of tannic acid (all final concentrations in PCR). The Applied Biosystems YFiler Plus PCR Amplification Kit was used for comparison.
Molecular biology grade distilled water
Manufactured by Thermo Fisher Scientific
Molecular biology grade distilled water is a purified water product designed for use in molecular biology applications. It is produced through a rigorous distillation process to ensure high purity and low levels of contaminants. This water is suitable for a variety of laboratory procedures where high-quality water is required.
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2 protocols using molecular biology grade distilled water
1
Evaluating YFiler Platinum Casework Kit Inhibition
2
Evaluating VeriFiler Plus Kit Performance Under PCR Inhibition
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VeriFiler Plus kit performance was tested in the presence of PCRinhibitory substances such as hematin, humic acid and tannic acid [17, 18] . In these experiments, samples composed of human male 007 genomic DNA containing different levels of PCR inhibitors were tested. Stock solutions of each inhibitor compound were prepared as follows: hematin (Sigma, St. Louis, MO) was dissolved in 0.1 N NaOH; humic acid (Alfa Aesar, Haverhill, MA) and tannic acid (Fisher Scientific, Pittsburgh, PA) were dissolved in molecular biology grade distilled water (Ambion, Austin, TX). Test samples were prepared with a constant input of 0.5 ng of 007 DNA per reaction but with increasing concentrations of each inhibitor: 400, 500 and 600 µM hematin, 150, 250 and 350 ng/µL of humic acid, or 250, 350 and 450 ng/µL of tannic acid (all final concentrations in PCR). Samples were amplified in replicates of four at each inhibitor level. The Applied Biosystems AmpFℓSTR™ Identifiler™ Plus Kit was used for comparison.
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