A comparison of methods for forensic DNA extraction: Chelex-100® and the QIAGEN DNA Investigator Kit (manual and automated)

Phillips, Kirsten; McCallum, Nicola; Welch, Lindsey

doi:10.1016/j.fsigen.2011.04.018

Cited by 77 publications

(52 citation statements)

References 2 publications

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“…The resin is composed of polystyrene divinylbenzene copolymers containing paired iminodiacetate ions, which act as chelators for polyvalent metal ions (see Table 1) 10 . This group can interact via its nitrogen and oxygen atoms with the mercury according a tridentate interaction.…”

Section: Methodsmentioning

confidence: 99%

Removal of Cd (II) and Hg(II) by chelating resin Chelex-100

Amara-Rekkab¹,

Didi²

2015

Orient. J. Chem

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A sensitive, simple method for the determination of amounts of mixture of Hg 2+ and Cd 2+ by spectrophotometry was described based on the formation of the Hg 2+ -Cd 2+ -PAN complex in water media. Optimal conditions such as reagent amounts, and pH for the Hg 2+ -Cd 2+ determination were reported. It was found that the 2:1 PAN-Hg 2+ -Cd 2+ complex dominate at pH 13.0. In another hand, the sorption of mixture mercury (II) and cadmium (II) from aqueous medium on a chelating resin Chelex 100 was studied in batch mode. Since the extraction kinetic was obtained, with a mixture of 0.1 g of resin and 5 mL of mixture at 1 mmol/L of initial concentration, extraction equilibrium was reached within 20 min of mixing. The influence of some parameters such as initial mixture ions concentrations, initial pH of aqueous solution, ion strength and the amounts of resin have been studied at fixed temperature (20±1°C). The optimum pH value level for quantitative sorption was up to 2.6. The best performance obtained was 97.1% of extraction yield equivalent to 15.65 mg/g of resin. The pseudo-first-order, pseudo-second-order models and the intra-particle diffusion model were used to describe the kinetic data and rate constants were evaluated.

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Section: Methodsmentioning

confidence: 99%

Removal of Cd (II) and Hg(II) by chelating resin Chelex-100

Amara-Rekkab¹,

Didi²

2015

Orient. J. Chem

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“…Several organic and inorganic protocols are available for DNA extraction. They vary in their nucleic acids yield, processing time, and the ability of removing the Polymerase Chain Reaction inhibitors (PCR inhibitors) (Phillips et al, 2012). DNA can be successfully extracted from a wide range of biological samples, these include, blood residues, urine, semen, saliva, soft and hard tissues.…”

Section: Introductionmentioning

confidence: 99%

Effects of storage temperature on the quantity and integrity of genomic DNA extracted from mice tissues: A comparison of recovery methods

Al-Griw¹,

Zraba²,

Al-Muntaser³

et al. 2017

Open Vet J.

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Efficient extraction of genomic DNA (gDNA) from biological materials found in harsh environments is the first step for successful forensic DNA profiling. This study aimed to evaluate two methods for DNA recovery from animal tissues (livers, muscles), focusing on the best storage temperature for DNA yield in term of quality, quantity, and integrity for use in several downstream molecular techniques. Six male Swiss albino mice were sacrificed, liver and muscle tissues (n=32) were then harvested and stored for one week in different temperatures, -20C, 4C, 25C and 40C. The conditioned animal tissues were used for DNA extraction by Chelex-100 method or NucleoSpin Blood and Tissue kit. The extracted gDNA was visualized on 1.5% agarose gel electrophoresis to determine the quality of gDNA and analysed spectrophotometrically to determine the DNA concentration and the purity. Both methods, Chelex-100 and NucleoSpin Blood and Tissue kit found to be appropriate for yielding high quantity of gDNA, with the Chelex100 method yielding a greater quantity (P < 0.045) than the kit. At -20C, 4C, and 25C temperatures, the concentration of DNA yield was numerically lower than at 40C. The NucleoSpin Blood and Tissue kit produced a higher (P=0.031) purity product than the Chelex-100 method, particularly for muscle tissues. The Chelex-100 method is cheap, fast, effective, and is a crucial tool for yielding DNA from animal tissues (livers, muscles) exposed to harsh environment with little limitations.

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“…La extracción del ADN, es el paso inicial en los procedimientos de un laboratorio de genética, consiste en aislar la molécula de ADN de cualquier tipo de célula nucleada, en condiciones de alta calidad, pureza y buena cantidad, que permita su caracterización genética (ChaconCortes et al 2012); se utilizan diferentes metodologías como el Chelex ® 100 y QIAGEN DNA Investigator Kit (Phillips et al 2012, Ps et al 1991) o el reactivo de purificación FTA (de Vargas Wolfgramm et al 2009, entre otros).…”

Section: Introductionunclassified

Variación de perfiles genéticos obtenidos por PCR con y sin extracción de ADN a partir de manchas de sangre

et al. 2017

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ResumenLa PCR sin extracción es una herramienta clave en investigaciones forenses, tratándose de una de las opciones a elegir cuando se tienen muestras con baja cantidad o calidad del ADN. El objetivo de este estudio fue determinar si hay diferencias entre los perfiles genéticos obtenidos por PCR i) en muestras de ADN extraídas con Chelex ® 100, ii) en muestras de ADN obtenidas con el reactivo de purificación de muestras en tarjetas FTA, y iii) por la PCR muestras sin extracción, empleando el kit comercial AmpFLSTR ® Identifiler ® , utilizado de rutina en el laboratorio IdentiGEN de la Universidad de Antioquia. Manchas de sangre almacenadas en tarjetas FTA TM Genecard Whatman TM , papel de Cromatografía y papel filtro, se amplificaron por PCR sin extracción utilizando los buffers Ezway TM , PCRboost TM , STRboost TM y Buffer 5X Colorless Go Taq ® Flexi. Los perfiles obtenidos se analizaron por medio de electroforesis capilar. Los resultados muestran que con los buffers utilizados se obtiene la amplificación de todos los marcadores STRs de las muestras almacenadas en los tres diferentes soportes. Esto permite concluir, que utilizando la PCR sin extracción, además de reducirse los costos y el tiempo de procesamiento de las muestras, se logra obtener perfiles genéticos que cumplan los criterios de calidad establecidos por el laboratorio, generando resultados de manera más eficiente al no ser necesario realizar la extracción y purificación del ADN.Palabras claves: PCR sin extracción, extracción de ADN, genética forense. AbstractThe PCR without extraction is a key tool in forensic investigation, being a good option when samples with small amounts or degraded DNA are available. The objective of this study was to compare genetic profiles obtained by PCR using: i) DNA samples extracted using Chelex ® 100, ii) DNA samples extracted using an FTA purification reagent, and iii) samples without extraction using the commercial kits AmpFLSTR ® Identifiler ® , routinely employed in the laboratory IdentiGEN of the University of Antioquia. Blood stains were collected in FTA cards Genecard Whatman TM , chromatography paper, and in filter papers. PCR without extraction was performed using the buffers Ezway TM , PCRboost TM , STRboost TM , and Buffer 5X Colorless Go Taq ® Flexi. The profiles were obtained by capillary electrophoresis. The results show that, using the above-mentioned buffers, all STR markers were amplified, for samples collected in the three different types of support. In conclusion, the PCR without extraction, apart from reducing the cost and time needed for processing the samples, it allows obtaining genetic profiles that meet the quality criteria established by the laboratory, in a more efficiently way, since DNA extraction and purification steps are eliminated.

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A comparison of methods for forensic DNA extraction: Chelex-100® and the QIAGEN DNA Investigator Kit (manual and automated)

Cited by 77 publications

References 2 publications

Removal of Cd (II) and Hg(II) by chelating resin Chelex-100

Removal of Cd (II) and Hg(II) by chelating resin Chelex-100

Effects of storage temperature on the quantity and integrity of genomic DNA extracted from mice tissues: A comparison of recovery methods

Variación de perfiles genéticos obtenidos por PCR con y sin extracción de ADN a partir de manchas de sangre

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