Evaluation of post-PCR purification methods and subsequent optimisation of the MinElute® PCR Purification Kit for low input DNA samples amplified with PowerPlex® 21.

Publisher: Elsevier Science Ireland Country of Publication: Ireland NLM ID: 7902034 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1872-6283 (Electronic) Linking ISSN: 03790738 NLM ISO Abbreviation: Forensic Sci Int Subsets: MEDLINE

Publication: Limerick : Elsevier Science Ireland
Original Publication: Lausanne, Elsevier Sequoia.

DNA Fingerprinting*/methods ; Polymerase Chain Reaction*/methods ; DNA*/isolation & purification ; Microsatellite Repeats*; Humans ; Alleles

Weak and partial DNA profiles are commonly encountered within forensic casework due to amplification of low DNA input samples. One option for increasing allelic detection in such samples is the purification of amplified PCR product using commercially available column-based methods. In this study, four commercially available post-PCR purification methods, QIAGEN MinElute®, Independent Forensics Amplicon™ Rx, Millipore Microcon® and Thermo Fisher Scientific ExoSAP-IT™ were evaluated, comparing the quality of PowerPlex® 21 DNA profiles produced to the standard DNA profile generated prior to purification. An increased detection of alleles above the analytical threshold was observed following purification with the MinElute®, Amplicon™ Rx and Microcon® methods, allowing informative DNA profiles to be recovered using as little as 8 pg DNA. However, post-PCR purification using the ExoSAP-IT™ kit was unsuccessful, with no alleles detected above analytical threshold in samples with ≤16 pg DNA. The MinElute® kit was selected for optimisation on the basis of DNA profile quality, including increased detection of alleles and minimal artefacts. The MinElute® method was optimised by evaluating the number of washes and final elution buffer volume, resulting in a further increase in detection of alleles by reducing the elution buffer volume. Overall, this study showed that PowerPlex® 21 DNA profiles from low input DNA can be successfully enhanced by employing the MinElute® post-PCR purification method.
Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Crown Copyright © 2024. Published by Elsevier B.V. All rights reserved.)

Keywords: Amplicon™ Rx; ExoSap-IT™; Low template DNA; Microcon® fast flow; MinElute®; Post-PCR Purification

9007-49-2 (DNA)

Date Created: 20240505 Date Completed: 20240609 Latest Revision: 20240609

20240610

10.1016/j.forsciint.2024.112043

38705055