Novel extraction chemistry and alternative amplification strategies for use with rootless hair shafts

Gutierrez, Ryan; LaRue, Bobby; Houston, Rachel

doi:10.1111/1556-4029.14763

Cited by 8 publications

(2 citation statements)

References 28 publications

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“…The differences between the Hair and Bone Kits include different binding and digest buffers, DTT is included only in the Hair kit, and different volumes and incubation times. A previous study confirmed that rootless hairs processed with the InnoXtract Hair method yielded probative autosomal STR and INNUL profiles [45,46].…”

Section: Introductionsupporting

confidence: 59%

“…Cold cases, such as a 1980 Newport Beach murder [29] and a 1977 southern California murder [30] were both solved through hair forensic DNA analysis, where the hair sample was the only evidence available. As forensic DNA testing expands to newer DNA typing techniques (microhaplotype [31], SNP [32,33], SAP [34], InDel [35,36], INNUL [37,38], NGS [39,40], etc.) and new analysis methods, such as forensic genetic genealogy, rootless hairs are increasingly becoming a more probative sample type [41].…”

Section: Introductionmentioning

confidence: 99%

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Validation of the InnoXtract™ DNA extraction method for bone, teeth, and rootless hair

Sinha

Brown

Holt

et al. 2023

Journal of Forensic Sciences

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Forensic casework samples often include human hairs, teeth, and bones. Hairs with roots are routinely processed for DNA analysis, while rootless hairs are either not tested or processed using mitochondrial DNA. Bones and teeth are submitted for human remains identifications for missing persons and mass disaster cases. DNA extraction from these low templates and degraded samples is challenging. The new InnoXtract DNA extraction method utilizes magnetic beads that are optimized to bind small DNA fragments, as small as 100 base pairs, to purify high‐yield DNA from compromised samples. This validation study evaluates InnoXtract's ability to obtain amplifiable DNA from samples such as rootless hairs and skeletal remains. Studies performed include sensitivity, stability, repeatability, reproducibility, non‐probative samples, and comparison to standard organic extractions. Sensitivity studies demonstrate average yield recoveries ranging from 53% to 100% and 73% to 85% for the InnoXtract hair and bone methods, respectively. Studies demonstrate consistent results across a range of sample types, such as insulted and un‐insulted bone and teeth, as well as hair shafts from donors of various ages, gender, race, and hair characteristics. The InnoXtract bone method outperformed organic extraction. The method was successfully automated on a MagMAX™ Express‐96, with recoveries over 70% relative to the manual version. InnoXtract has the potential as an automated high‐throughput, high‐yield bone extraction method with 6 h of total extraction time for up to 96 samples. The validation study results demonstrate that the InnoXtract kits produce high‐yield and high‐quality DNA from compromised bone, teeth, and hair shaft samples.

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Section: Introductionsupporting

confidence: 59%

Section: Introductionmentioning

confidence: 99%

Validation of the InnoXtract™ DNA extraction method for bone, teeth, and rootless hair

Sinha

Brown

Holt

et al. 2023

Journal of Forensic Sciences

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Deoxyribonucleic Acid Extraction Methods in Forensic Analysis

Fernandez-Tejero¹,

Ghemrawi²,

McCord³

et al. 2022

Encyclopedia of Analytical Chemistry

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Forensic biological evidence encompasses a diverse conglomerate of samples containing genetic material found in a variety of biological fluids and tissues such as human blood, semen, saliva, epithelial cells, hair, bone, teeth, fingernails, and putrefied tissues. In addition to human samples, nonhuman samples from plant, animal, bacteria, and fungi may also need to be processed. The samples and body fluids may be present as dried stains on an assortment of substrates, often mixed with polymerase chain reaction (PCR) inhibitors, exposed to environmental conditions and uncontrolled degradation, and are often present in limited quantities. In addition to the intrinsic characteristics of samples that include the type of body fluid (e.g. blood, saliva, semen), nature of the tissues (e.g. buccal cells, hair, bone, tooth), quantity [e.g. trace DNA (deoxyribonucleic acid), low copy number], and source (e.g. reference, evidence), they can also be grouped by their extrinsic characteristics such as the substrate (e.g. swab, clothing) and nature of the crime (e.g. sexual assault, burglary, homicide) from which they are derived. These samples need to be processed using the most effective methods of nucleic acid extraction and purification for downstream quantification and genetic profiling by PCR. Compositionally, there are an unlimited number of combinations of sample and substrate types including the quantity and quality of the sample, substrate and conditions encountered, and contaminant and inhibitor levels. Some factors may be observed during screening, whereas many of these remain transparent to the analyst. This renders the choice of the most efficient DNA extraction methods, one of the most unpredictable steps in forensic DNA profiling. Research has led to significant enhancements, testing, and validation of advanced extraction and amplification chemistries along with new instrumentation platforms providing systems capable of automated, sensitive, rapid, and robust extraction and genotyping and/or sequencing of short tandem repeats (STR), single‐nucleotide polymorphisms (SNPs), and mitochondrial DNA. All three genetic marker classes have been accepted in the court systems worldwide and this along with legislation that has expanded both the law and funding for forensic DNA profiling has led to a high and increasing demand for forensic DNA testing. The increased demand and newly expanded capabilities have opened the door to the analysis of forensic samples that are extremely compromised in both quality and quantity. Over the past 10 years, new DNA extraction methods have been developed to reproducibly extract DNA and remove or mitigate PCR inhibitors from the majority of sample types with speed and efficiency, resulting in high‐yield template free of PCR inhibition. In addition, automated processing has been tested and validated to meet the demand for high‐throughput processing. This article summarizes the importance of DNA extraction, provides an update on methods for single source, compromised evidence, sexual assault evidence, automation, differential extraction, and an overview of the most widely adopted methods and the best practices for DNA isolation from forensic biological samples.

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DNA and protein analyses of hair in forensic genetics

Simayijiang

Wang

et al. 2023

Int J Legal Med

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Novel extraction chemistry and alternative amplification strategies for use with rootless hair shafts

Cited by 8 publications

References 28 publications

Validation of the InnoXtract™ DNA extraction method for bone, teeth, and rootless hair

Validation of the InnoXtract™ DNA extraction method for bone, teeth, and rootless hair

Deoxyribonucleic Acid Extraction Methods in Forensic Analysis

DNA and protein analyses of hair in forensic genetics

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