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7. Troubleshooting................................................................................................................................ 49
7.A. Amplification and Fragment Detection ....................................................................................... 49
7.B. Amplification of Extracted DNA ................................................................................................. 52
7.C. Direct Amplification of DNA From Storage Card Punches ............................................................ 53
7.D. Direct Amplification of DNA From Swabs ................................................................................... 56
7.E. GeneMapper® ID-X Software .................................................................................................... 59
7.F. GeneMapper® ID Software ........................................................................................................ 61
8. References ........................................................................................................................................ 64
9. Appendix .......................................................................................................................................... 65
9.A. Advantages of Using the Loci in the PowerPlex® Fusion System .................................................... 65
9.B. DNA Extraction and Quantification Methods and Automation Support ......................................... 69
9.C. The CC5 Internal Lane Standard 500 ......................................................................................... 70
9.D. Composition of Buffers and Solutions ......................................................................................... 70
9.E. Related Products ...................................................................................................................... 71
9.F. Summary of Changes................................................................................................................. 72
1.
Description
STR (short tandem repeat) loci consist of short, repetitive sequence elements 3–7 base pairs in length (1–4). These
repeats are well distributed throughout the human genome and are a rich source of highly polymorphic markers, which
may be detected using the polymerase chain reaction (5–9). Alleles of STR loci are differentiated by the number of
copies of the repeat sequence contained within the amplified region and are distinguished from one another using
fluorescence detection following electrophoretic separation.
The PowerPlex® Fusion System(a–g) is a 24-locus multiplex for human identification applications including forensic
analysis, relationship testing and research use. This five-color system allows co-amplification and fluorescent detection
of the 13 core CODIS (US) loci (CSF1PO, FGA, TH01, TPOX, vWA, D3S1358, D5S818, D7S820, D8S1179, D13S317,
D16S539, D18S51 and D21S11), the 12 core European Standard Set loci (TH01, vWA, FGA, D21S11, D3S1358,
D8S1179, D18S51, D10S1248, D22S1045, D2S441, D1S1656 and D12S391) and Amelogenin for gender determination.
In addition, the male-specific DYS391 locus is included to identify null Y allele results for Amelogenin. The Penta D
and Penta E loci are included to increase discrimination and allow searching of databases that include profiles with
these Penta loci. Finally, the D2S1338 and D19S433 loci, which are popular loci included in a number of databases,
were incorporated to further increase the power of discrimination. This extended panel of STR markers is intended to
satisfy both CODIS and ESS recommendations.
The PowerPlex® Fusion System is compatible with the ABI PRISM® 3100 and 3100-Avant Genetic Analyzers and
Applied Biosystems® 3130, 3130xl, 3500 and 3500xL Genetic Analyzers. Amplification and detection instrumentation
may vary. You may need to optimize protocols including amount of template DNA, cycle number, injection conditions
and loading volume for your laboratory instrumentation. In-house validation should be performed. We have tested the
PowerPlex® Fusion System with Data Collection Software, Versions 2.0 and 3.0, and GeneMapper® ID-X Software,
Versions 1.2, and 3.2. Other software versions may be available for use; however, the options available in other
versions may differ slightly from the options listed in this Technical Manual.
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TMD039 · Revised 3/15
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