Initial Evaluation of A96-Plex Goldengate® Genotyping SNP Assay with Suboptimal and Whole Genome Amplified Samples
Journal Title: Journal of Forensic Investigation - Year 2013, Vol 1, Issue 2
Abstract
A custom designed 96-plex GoldenGate® Genotyping single nucleotide polymorphism (SNP) assay was evaluated for performance with genomic samples (10-250ng template), whole genomic amplified (WGA) samples and environmentally challenged samples. The assay performed well with pristine genomic samples, reproducibly generating complete and accurate SNP profiles with lower (50ng) than the manufacturer’s recommended amount of template (250ng). Clinical and forensic samples often fall below the optimal concentration for direct SNP analysis. One proposed solution to this is to produce sufficient quantities of DNA prior to SNP typing by whole genome amplification (WGA). The results of this study show that WGA prior to SNP typing produced reliable results when the template was of high quality and quantity (≥10ng). However, SNP-typing of environmentally challenged skeletal samples produced poor results, both with and without WGA prior to SNP typing. The amplification bias inherent in the WGA process was significantly exaggerated with samples of low quality and quantity. While these results suggest that SNP-typing using the Illumina GoldenGate® assay is not the solution for genotyping highly degraded samples, it can provide an alternative means for some forensic DNA typing needs, such as paternity testing or typing reference samples for databasing.
Authors and Affiliations
Sheree Hughes-Stamm
Keywords
Related Articles
- EP ID EP202457
- DOI 10.13188/2330-0396.1000005
- Views 99
- Downloads 0
Forensic Evidence and Crime Scene Investigation
Contemporary law enforcement has greatly expanded its ability to solve crimes by the adoption of forensic techniques and procedures [1]. Today, crimes often can be solved by detailed examination of the crime scene and an...
A Comparison of Maceration Techniques for Use in Forensic Skeletal Preparations
Maceration removes all soft tissue from the bones. It must be done without damaging or morphing the bone. To examine the effectiveness of methods, six techniques were applied to a convenience sample of skeletal elements:...
Kinship Analysis Based on SNP Data from the HID-Ion AmpliSeqTM Identity Panel
Next generation sequencing supports extensive single nucleotide polymorphism (SNP) analysis owing to its comprehensive sequencing capacity. In the present study, a number of 20 parent–child pairs and 17 full siblings wer...
A Multipurpose Composition Based on Brilliant Blue R for Developing Latent Fingerprints on Crime Scene Evidence
The simplest and most commonly used method for detecting latent fingerprints is powder dusting. This technique relies on the mechanical adherence of the detecting composition to the moisture content of sweat. Under the p...
A New DFO Method for Developing Latent Fingermarks on Thermal Paper
A new method for improvement of DFO-treated latent fingermarks on thermal paper will be described in this paper. A DFO powder suspension was prepared using an ultrasonic cleaner to ensure DFO’s maximum solubility. The ne...