Highlights
- •Normalized extraction is combined with fast PCR and POP-6/22 cm array CE detection.
- •This process utilizes PowerPlex 16 HS, Identifiler Plus and Identifiler primer sets.
- •Processing time – from accessioning to analysis – is reduced by ∼37%.
- •High quality STR profiles were obtained from 95 to 99% of buccal swab samples.
- •High quality STR profiles were obtained from 88 to 91% of buccal collector punches.
Abstract
The goal of this project was to develop a “normalized” extraction procedure to be
used in conjunction with previously validated 3 μL fast PCR reactions (42–51 min utilizing KAPA2G™ Fast Multiplex PCR Kit) and alternative capillary electrophoresis
(24–28 min injection using POP-6™ Polymer and a 22 cm array). This was the final phase of a workflow overhaul for the database unit at
Cellmark Forensics to achieve a substantial reduction in processing time for forensic
DNA database samples without incurring significant added costs and/or the need for
new instrumentation, while still generating high quality STR profiles. Extraction
normalization aimed to consistently yield a small range of DNA concentrations, thereby
eliminating the need for sample quantification and dilution. This was specifically
achieved using the ChargeSwitch® Forensic DNA Purification Kit and a reduction in
extraction bead quantity, thereby forcing an increase in bead binding efficiency.
Following development of this extraction procedure, an evaluation ensued to assess
the combination of normalized extraction, 3 μL fast PCR (with PowerPlex 16 HS, Identifiler Plus and Identifiler primer sets),
and alternative CE detection – further referred to as new “first pass” procedures.
These modifications resulted in a 37% reduction in processing time and were evaluated
via an in depth validation, from which nearly 2000 STR profiles were generated, of
which 554 profiles from 77 swab donors and 210 profiles from 35 buccal collector donors
specifically arose from the new first pass procedures. This validation demonstrates
the robustness of these processes for buccal swabs and Buccal DNA Collectors™ using
the three primer sets evaluated and their ability to generate high quality STR profiles
with 95–99% and 88–91% pass rates, respectively.
Keywords
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Article info
Publication history
Published online: August 03, 2016
Accepted:
July 28,
2016
Received in revised form:
May 27,
2016
Received:
March 10,
2016
Identification
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© 2016 Elsevier Ireland Ltd. All rights reserved.
