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tmd017.0708.qxp
7/25/2008
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Appendix ...................................................................................................................................44
A. Advantages of STR Typing...........................................................................................44
B. Advantages of Using the Loci in the PowerPlex® ES System .................................44
C. DNA Extraction and Quantitation Methods..............................................................47
D. The Internal Lane Standard 600...................................................................................47
E. PowerPlex® ES Separation Control .............................................................................48
F.
Preparing the PowerPlex® ES System Master Mix ...................................................48
G. Composition of Buffers and Solutions........................................................................49
H. Related Products ............................................................................................................49
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 (5–8). 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 radioactive, silver stain or fluorescence
detection following electrophoretic separation.
The PowerPlex® ES System(a–c) allows co-amplification and three-color detection of
nine loci (eight STR loci and Amelogenin), including D18S51, D21S11, TH01,
D3S1358, FGA, D8S1179, vWA, Amelogenin and SE33. One primer specific for each
of the D18S51, D21S11, TH01 and D3S1358 loci is labeled with fluorescein (FL); one
primer specific for each of the FGA, D8S1179, vWA and Amelogenin loci is labeled
with carboxy-tetramethylrhodamine (TMR); and one primer specific for SE33 is
labeled with 6-carboxy-4´,5´-dichloro-2´,7´-dimethoxy-fluorescein (JOE). All nine loci
are amplified simultaneously in a single tube and analyzed in a single injection.
The PowerPlex® ES Monoplex System, SE33 (JOE) (Cat.# DC6751) is available to
amplify the SE33 locus. This monoplex system allows amplification of a single locus
to confirm results obtained with the PowerPlex® ES System and re-amplify DNA
samples when one or more of the loci do not amplify initially due to nonoptimal
amplification conditions or poor DNA template quality.
The PowerPlex® ES System is compatible with the ABI PRISM® 310, 3100 and 3100Avant Genetic Analyzers and Applied Biosystems 3130 and 3130xl Genetic Analyzers.
The protocols presented in this manual were tested at Promega Corporation.
Amplification and detection instrumentation may vary. You may need to optimize
protocols including cycle number and injection time (or loading volume) for each
laboratory instrument. In-house validation should be performed.
Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · Phone 608-274-4330 · Fax 608-277-2516 · www.promega.com
Part# TMD017
Page 2
Printed in USA.
Revised 7/08