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Assessment of DNA Extracted from Forensic Samples Prior to Genotyping.

M Barbisin¹, J G Shewale¹

¹Life Technologies Corporation, Foster City, CA, USA.

Forensic Science Review

|August 6, 2015

Summary

Real-time PCR methods now quantify human DNA and assess DNA quality for forensic analysis. This improves genotyping success rates, especially with limited evidence samples.

Keywords:

Assessment of DNA extract DNA analysis DNA quantification DNA typing assessment of forensic samples human DNA human male DNA real-time PCR

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Area of Science:

Forensic Science
Molecular Biology
Genetics

Background:

Early forensic DNA analysis relied on hybridization-based methods like Quantiblot® kits, suitable for 1990s genotyping technologies.
Advancements in genotyping, including short tandem repeat (STR) profiling, mitochondrial DNA (mtDNA) sequencing, and single nucleotide polymorphism (SNP) typing, necessitated more sensitive DNA quantification and quality assessment.

Purpose of the Study:

To address the need for picogram-level DNA quantification and quality assessment in modern forensic science.
To highlight the utility of real-time PCR methods for optimizing downstream genotyping success rates.

Main Methods:

Development and application of real-time PCR (polymerase chain reaction) assays for DNA quantification.
Singleplex and multiplex real-time PCR systems capable of quantifying human DNA, human male DNA, mtDNA, detecting PCR inhibitors, and assessing DNA degradation.

Main Results:

Real-time PCR methods provide sensitive quantification (picogram level) and crucial quality assessment of DNA extracts.
Multiplex real-time PCR assays can analyze multiple targets simultaneously, enhancing efficiency.
Integration of these assays improves the selection of genotyping systems and increases first-pass success rates.

Conclusions:

Real-time PCR-based quantification and quality assessment are essential for modern forensic DNA analysis.
These methods enable informed decisions, maximizing the success of genotyping from minimal evidence samples.
The described real-time PCR assays serve as valuable DNA assessment tools for various genotyping and sequencing applications in human identification.