Download PowerPlex® 16 System

tmd012.0508.qxp
9/16/2008
3:47 PM
IX.
I.
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Appendix ...................................................................................................................................46
A. Advantages of STR Typing...........................................................................................46
B. Advantages of Using the Loci in the PowerPlex® 16 System..................................47
C. Power of Discrimination ...............................................................................................50
D. DNA Extraction and Quantitation Methods..............................................................52
E. The Internal Lane Standard 600...................................................................................52
F.
Preparing the PowerPlex® 16 System Master Mix ....................................................53
G. Agarose Gel Electrophoresis of Amplification Products (Optional)......................54
H. Composition of Buffers and Solutions........................................................................55
I.
Related Products ............................................................................................................56
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® 16 System(a–d) (9,10) allows co-amplification and three-color
detection of sixteen loci (fifteen STR loci and Amelogenin), including Penta E,
D18S51, D21S11, TH01, D3S1358, FGA, TPOX, D8S1179, vWA, Amelogenin, Penta D,
CSF1PO, D16S539, D7S820, D13S317 and D5S818. One primer for each of the Penta E,
D18S51, D21S11, TH01 and D3S1358 loci is labeled with fluorescein (FL); one primer
for each of the FGA, TPOX, D8S1179, vWA and Amelogenin loci is labeled with
carboxy-tetramethylrhodamine (TMR); and one primer for each of the Penta D,
CSF1PO, D16S539, D7S820, D13S317 and D5S818 loci is labeled with 6-carboxy-4´,5´dichloro-2´,7´-dimethoxy-fluorescein (JOE). All sixteen loci are amplified
simultaneously in a single tube and analyzed in a single injection or gel lane.
The PowerPlex® 16 Monoplex System, Penta E (Fluorescein) (Cat.# DC6591) and
PowerPlex® 16 Monoplex System, Penta D (JOE) (Cat.# DC6651) are available to
amplify the Penta E and Penta D loci, respectively. Each monoplex system allows
amplification of a single locus to confirm results obtained with the PowerPlex® 16
System, PowerPlex® 16 BIO System or PowerPlex® 2.1 System. The monoplex
systems can be also used to 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® 16 System is compatible with the ABI PRISM® 310, 3100 and 3100Avant Genetic Analyzers, Applied Biosystems 3130 and 3130xl Genetic Analyzers
and ABI PRISM® 377 DNA Sequencer. 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# TMD012
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Printed in USA.
Revised 5/08