Higher Capillary Electrophoresis Injection Settings as an Efficient Approach to Increase the Sensitivity of STR Typing

• Published in: Journal of forensic sciences Vol. 54; no. 3; pp. 591 - 598
• Main Authors: Westen, Antoinette A., Nagel, Jord H. A., Benschop, Corina C. G., Weiler, Natalie E. C., De Jong, Bas J., Sijen, Titia
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01-05-2009; Wiley Subscription Services, Inc
• Subjects: admixed samples; Alleles; capillary electrophoresis; DNA - analysis; DNA Fingerprinting - methods; DNA fingerprints; DNA typing; Electrophoresis, Capillary - methods; forensic science; Forensic sciences; Gene amplification; Genes; Genotype; Humans; low copy number; low-level DNA; Polymerase Chain Reaction; Studies; Tandem Repeat Sequences
• ISSN: 0022-1198; 1556-4029
• DOI: 10.1111/j.1556-4029.2009.01022.x

:  Evidentiary traces may contain low quantities of DNA, and regularly incomplete short tandem repeat (STR) profiles are obtained. In this study, higher capillary electrophoresis injection settings were used to efficiently improve incomplete STR profiles generated from low‐level DNA samples under standard polymerase chain reaction (PCR) conditions. The method involves capillary electrophoresis with higher injection voltage and extended injection time. STR peak heights increased six‐fold. Inherent to the analysis of low‐level DNA samples, we observed stochastic amplification artifacts, mainly in the form of allele dropout and heterozygous peak imbalance. Increased stutter ratios and allele drop‐in were rarely seen. Upon STR typing of 10:1 admixed samples, the profile of the major component did not become overloaded when using higher injection settings as was observed upon elevated cycling. Thereby an improved profile of the minor component was obtained. For low‐level DNA casework samples, we adhere to independent replication of the PCR amplification and boosted capillary electrophoresis.