Ann M. Lins¹, Steve Creacy², Cynthia J. Sprecher¹, Katherine A. Micka¹, Dawn R. Rabbach¹, Robert A. Bever², and James W. Schumm^1
1Promega Corporation, Madison, WI
²Genetic Design, Greensboro, NC

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We have developed a STR multiplex amplification system which allows simultaneous amplification of 8 STR loci. This single tube system generates alleles of four loci labeled with flurorescein (i.e. CSF1PO, TPOX, TH01, vWA) and four more labeled with carboxy-tetramethylrhodamine (TMR) (i.e. D16S539, D7S820, D13S317, D5S8180). Alleles from each locus labeled with the same dye do not overlap with neighboring loci. The inclusion of allelic ladders with the system provides a rapid and precise method for allele determination.

All eight loci may be detected in a single gel lane using either the high throughput approach of the Hitachi FMBIO^® Fluorescent Scanner or the moderate throughput systems of the Applied Biosystems 373 and Prism^tm 377 DNA Sequencers.

We have characterized this system for the following properties:

• Template sensitivity – comparison of amplifications in the Perkin Elmer 480 and GeneAmp 9600 thermal cyclers.
• Comparison of locus to locus peak heights – comparison of amplifications in the Perkin Elmer 480
  and the GeneAmp 9600 thermal cyclers.
• Effects of modifying annealing temperature from the recommended amplification protocol
  results using AmpliTaq^tm or AmpliTaq^tm Gold.

In addition, a population study of the loci making up the GenePrint^™ PowerPlex^™ System has been completed. The Matching Probability, Typical Paternity Index, and Calculated Power of Exclusion have been determined for the entire system in each of three population groups as shown in table below.

These data indicate that the GenePrint^™ PowerPlex^™ System is an ideal system for efficient generation of databases. In addition, the strong Typical Paternity Indices displayed by this single system are appropriate for use in paternity determination.