A novel, 4-h DNA extraction method for STR typing of casework bone samples

Hasap, Laila; Chotigeat, Wilaiwan; Pradutkanchana, Jintana; Vongvatcharanon, Uraporn; Kitpipit, Thitika; Thanakiatkrai, Phuvadol

doi:10.1007/s00414-019-02232-9

Cited by 17 publications

(6 citation statements)

References 52 publications

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“…The concentration and pH of EDTA were previously reported to affect the demineralization process of the teeth or on the bovine cortical bone (Sales et al 2018). Hasap et al (2020) found that total demineralization combined with DNA purification by commercial kit improved DNA yield and STR profiles (Hasap et al 2020). However, most tested bone samples would be around the age between less than 1 to 4 years and few samples would be around the ages 5 to 7 years, whereas this research used more than 20 years old preservation of skeletal remains.…”

Section: Discussionmentioning

confidence: 99%

“…A highly efficient DNA extraction method has been developed such as total demineralization plus silica membrane columns method or modified Hi-Flow® silica method, and PrepFiler® BTA kit. Total demineralization of bone powder significantly increased DNA yield and improve STR typing results (Lee et al 2010;Seo et al 2010;Hasap et al 2020). Although the concentration of EDTA in the demineralization process has been reported to affect DNA extraction, other factors including pH of EDTA solution, incubation temperature, incubation time, and volume of EDTA solution have been probably suggested to affect complete total demineralization.…”

Section: Introductionmentioning

confidence: 99%

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Factors influencing DNA extraction from human skeletal remains: bone characteristic and total demineralization process

Booncharoen

Khacha-ananda

Kanchai

et al. 2021

Egypt J Forensic Sci

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Background The extraction of DNA from skeletal remains with good quality and quantity is often challenging for the ability to generate DNA typing. Previous studies demonstrated the DNA extraction with total demineralization from fresh teeth and bones; however, the application in old skeletal remains has been less performed. To obtain good quality and high yield of DNA amount extracted from skeletal remains, the objective of this study was focused on exploring the factors influencing the total demineralization process to obtain developing effective methods. Results The concentration of EDTA was found to significantly enhance calcium chelation from the bone while pH of EDTA solution, incubation temperature, incubation time, and volume of EDTA solution were not significant. The optimal condition of total demineralization obtained from Placket-Burmann results represented good-quality DNA and the highest concentration of extracted DNA yield. Subsequently, the STR typing in some bone specimens processed by total demineralization process prior to DNA extraction was improved. Conclusions EDTA concentration was a key influencing factor on the total demineralization process to chelate calcium from human skeletal remains. The total demineralization process in old bone specimens probably improved the STR profiles.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Factors influencing DNA extraction from human skeletal remains: bone characteristic and total demineralization process

Booncharoen

Khacha-ananda

Kanchai

et al. 2021

Egypt J Forensic Sci

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“…Since this research was conducted, another study has shown that PrepFiler™ BTA is often sufficient for genotyping degraded bones after demineralisation of large bone fragments [49]. Additionally, Hasap et al [50] describe a novel 4 h DNA extraction method for STR typing of casework bone samples. Our protocols are faster, combining and modifying previous work [22,23] to develop a powderfree, rapid 15 min PrepFiler™ Lysis Buffer incubation that is compatible with downstream automated processing; reducing sample preparation and extraction to 2 h. Applying minimally-invasive samples such as nail and distal phalanges further expedites the process.…”

Section: Novelty and Practicalitymentioning

confidence: 99%

Efficient DNA Profiling Protocols for Disaster Victim Identification

et al. 2021

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Advances in forensic biology have increased the options for the collection, sampling, preservation and processing of human remains for DNA-based identification. Combined with a plethora of commercial DNA testing kits that are far more forgiving of inhibited and degraded samples, efficient DNA approaches to post-mortem samples are explored here for DNA-based identification of compromised human remains. Approaches which preserve sample and reduce analytical turnaround times whilst saving resources also have the potential to expedite the identification process, to provide answers to grieving families sooner, or to provide leads in a criminal investigation. Targeting sample types that are minimally-invasive and do not require extensive preparation and testing protocols also has benefit for disaster victim identification (DVI) by facilitating field sampling. We have assessed minimally-invasive and simple to collect sample types compatible with minimal pre-treatment and efficient DNA profiling approaches. Incubating nail, distal phalanges and whole digits in 500 µL of PrepFiler™ Lysis Buffer for 2 h was an efficient and simple method, limiting or removing sample preparation. A reduced 15 min incubation also yielded DNA profiles suggesting a shorter incubation may lyse sufficient DNA. Preservative solutions offer an even simpler process in some cases. Furthermore, the efficient approaches described in this study offer storage solutions and are compatible with backend automated processing. This study will inform further research to develop and optimise efficient protocols. These DNA approaches should not be pursued for every sample; more compromised samples may best be submitted to the laboratory for more effective extraction and genotyping.

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“…To ensure comparability of the results, all other parameters in the experiment were standardized during the analyses (Supplementary Material 1). Recently, Hasap et al published an improved method for forensic casework samples which proposes a lysis time of only 2 h [13]. We therefore considered this significant reduction in lysis time even for historical samples.…”

Section: Introductionmentioning

confidence: 99%

The Influence of Sample Quantity and Lysis Parameters on the Success of Ancient DNA Extraction from Skeletal Remains

et al. 2021

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DNA extraction is of utmost importance in archaeobiology, as it determines the success of further DNA analyses. This study concentrates on the success of ancient DNA extraction using silica spin columns and PCR-based analysis from archaeological skeletal material and investigates the influence of sample quantity, lysis time and lysis temperature during sample preparation. The results show that lysis times ranging from 2 to 48 h are suitable, and that lysis should be carried out at a constant temperature of 56°C. Concerning sample quantity, 10 mg for mitochondrial DNA and 50 mg for chromosomal DNA are sufficient for high quality analyses. Thus invaluable sample material can be saved, and time of sample preparation can be reduced considerably.

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A novel, 4-h DNA extraction method for STR typing of casework bone samples

Cited by 17 publications

References 52 publications

Factors influencing DNA extraction from human skeletal remains: bone characteristic and total demineralization process

Factors influencing DNA extraction from human skeletal remains: bone characteristic and total demineralization process

Efficient DNA Profiling Protocols for Disaster Victim Identification

The Influence of Sample Quantity and Lysis Parameters on the Success of Ancient DNA Extraction from Skeletal Remains

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