Download PowerPlex® Y23 System

7. Troubleshooting................................................................................................................................ 48
7.A. Amplification and Fragment Detection ....................................................................................... 48
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 .......................................................................................................................................... 66
9.A. Advantages of Using the Loci in the PowerPlex® Y23 System ........................................................ 66
9.B. Detection of Amplified Fragments Using the Applied Biosystems® 3130 or 3130xl
Genetic Analyzer with POP-7™ Polymer and Data Collection Software, Version 3.0 ....................... 70
9.C DNA Extraction and Quantification Methods and Automation Support ......................................... 73
9.D. The CC5 Internal Lane Standard 500 Y23 ................................................................................... 74
9.E. Composition of Buffers and Solutions ......................................................................................... 74
9.F. Related Products ...................................................................................................................... 75
9.G. Summary of Changes................................................................................................................. 76
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.
STR markers on the Y chromosome (Y-STR) have qualities that are distinct from autosomal markers and are useful for
human identification (10–16). Y-STR markers are found on the nonrecombining region of the Y chromosome (NRY) and
produce a haploid profile when amplified from male DNA. This quality simplifies male/female mixture interpretation
by removing the female contribution from an amplification profile (17,18). Strict paternal inheritance of these markers
makes them useful for paternity and kinship studies.
The PowerPlex® Y23 System(a–d) allows co-amplification and four-color fluorescent detection of 23 loci, including
DYS576, DYS389I, DYS448, DYS389II, DYS19, DYS391, DYS481, DYS549, DYS533, DYS438, DYS437, DYS570,
DYS635, DYS390, DYS439, DYS392, DYS643, DYS393, DYS458, DYS385a/b, DYS456 and Y-GATA-H4.
The PowerPlex® Y23 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 the 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® Y23 System with Data Collection Software, Versions 2.0 and 3.0, 3500 Data Collection Software Version 1.0,
GeneMapper® ID-X Software, Version 1.2, and GeneMapper® ID Software, Version 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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TMD035 · Revised 3/15
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