High Throughput Manual STR Analyses

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High Throughput Manual STR Analyses

Dawn R. Rabbach,¹, Katherine A. Micka,¹ Cynthia J. Sprecher,¹ Ann M. Lins¹, Jeffery W. Bacher¹, Steve Creacy²,
Robert A. Bever², and James W. Schumm¹
¹Promega Corporation, Madison, WI, USA
²Genetic Design, Inc., Greensboro, NC, USA.

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STR loci provide an efficient, rapid, and accurate method of typing DNA samples. We have developed a set of multiplex STR systems which are ideal for database and casework analysis. They include the following features:

High discrimination power.

Loci have been selected to minimize artifacts such as stutter bands and microvariants.

Multiplex STR loci may be analyzed simultaneously.

There is no overlap between the allele sizes of STR loci detected with the same dye or stain.

There is balance between the alleles within and between each locus.

Allelic ladders are provided with each system to simplify and add precision.

Three multiplex systems compatible with silver detection have been defined (1) GenePrint Ō CTT Multiplex System¹ (CSF1PO, TPOX, TH01), the GenePrint Ō FFv Multiplex System¹ (F13A01, FESFPS, vWA), and the GenePrint Ō SilverSTRŌ III Multiplex System (D16S539, D7S820, D13S317).

There is locus compatibility across manual and instrument-based detection formats. The new GenePrint Ō SilverSTRŌ III Multiplex System (D16S539, D7S820, D13S317), and the GenePrint Ō CTT Multiplex System¹ (CSF1PO, TPOX, TH01), contain six of the eight loci found in the GenePrintŌ PowerPlexŌ System. The GenePrint Ō FFv Multiplex System¹ (F13A01, FESFPS, vWA) contains the vWA locus also found in the GenePrintŌ PowerPlexŌ System. Therefore, seven of the eight loci found in the GenePrintŌ PowerPlexŌ System are found in our GenePrint Ō Silver Triplex Systems. Accordingly, once a laboratory has decided to invest in an instrument-based detection format, there is easy conversion to the GenePrintŌ PowerPlexŌ System because of the prior familiarity with the manual detection systems.

Analysis of over 200 individuals in each of three racial/ethnic groups indicates that using the three silver multiplexes the Matching Probability for these nine loci exceeds 1 in 1,000,000,000 for each racial group tested. Each of the silver stain systems may be evaluated without the need for expensive instrumentation. Use of these systems is very inexpensive because it requires only standard electrophoresis equipment and no specialized detection instruments.

*All 3 GenePrintŌ Silver STR Systems* Multiplexes (9 STR Loci)	African-American	Caucasian-American	Hispanic-American
Matching Probability	1 in 4.93 x 10⁹	1 in 1.05 x 10⁹	1 in 1.83 x 10⁹
Typical Paternity Index	987	723	560
Power of Exclusion	.9991	.9989	.9986

^{1 Multiplex Sets for Amplification of Polymorphic Short Tandem Repeat Loci --
Silver Stain and Fluorescent Detection, (1996) Lins, A.M., Sprecher, C.J., Puers, C., and
Schumm, J.W. Biotechniques 20: 882-889.}

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