Characterization and Validation Studies of Powerplex™ 2.1, A Nine-locus Short Tandem Repeat (Str) Multiplex System and Penta D Monoplex

Levedakou, Eleni N.; Freeman, David A.; Budzynski, Michael J.; Early, Buddy E.; Damaso, Ruth C; Pollard, Anne M; Townley, Amy Jo; Gombos, Jennifer L; Lewis, Jennifer L; Kist, Frank; Hockensmith, Mary E; Terwilliger, Michelle L; Amiott, Elizabeth A.; McElfresh, Kevin C.; Schumm, James W.; Ulery, Suzanna R; Konotop, Felipe; Sessa, Tara; Sailus, Jeffrey S; Crouse, Cecelia A.; Tomsey, Christine S.; Ban, Jeffrey D.; Nelson, Mark

doi:10.1520/jfs15443j

Cited by 15 publications

(6 citation statements)

References 25 publications

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“…In addition, amplicons were confined to individual loci and, therefore, no characteristic profiles could be obtained. Similar observations have recently been described for several other multiplex systems (Sparkes et al 1996;Wallin et al 1998;LaFountain et al 2001;Levedakou et al 2002). At least for primates, the appearance of these amplification products may be attributed to a partial homology within the primer binding regions of the same loci (Wallin et al 1998).…”

Section: Species Specificitysupporting

confidence: 84%

Validation of the multiplex kit genRES MPX-2 for forensic casework analysis

Junge¹,

Lederer

Braunschweiger³

et al. 2003

International Journal of Legal Medicine

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Validation studies were carried out using the commercially available PCR multiplex system genRESMPX-2. In addition to amelogenin, this system comprises the complete set of eight STR systems which are components of the German DNA database established in 1998 by the Federal Criminal Office of Germany (BKA). The minimum amount of template DNA which gave a complete DNA pattern ranged between 100 pg and 200 pg. Mixed samples could clearly be assigned from ratios between 1:5 (ACTBP2) and 1:20 (VWA, FGA). Experimental investigations with different forensic materials, environmental studies, reproducibility and precision data as well as practical casework analysis revealed that the genRESMPX-2 kit can be regarded as a sensitive, reliable and robust multiplex system even in the case of samples containing limited amounts or degraded DNA.

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Section: Species Specificitysupporting

confidence: 84%

Validation of the multiplex kit genRES MPX-2 for forensic casework analysis

Junge¹,

Lederer

Braunschweiger³

et al. 2003

International Journal of Legal Medicine

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“…In the late 1990s, Applied Biosystems began providing the Profiler Plus and COfiler kits to enable coverage of the 13 core loci for use on their instrument platforms (22). Promega Corporation developed the PowerPlex 2.1 kit to cover the additional loci not present in their PowerPlex 1.1 kit for use on the FMBIO detection platform (1,20,23). Table 1 summarizes the various STR kits that have become available in the past decade.…”

mentioning

confidence: 99%

Genetics and Genomics of Core Short Tandem Repeat Loci Used in Human Identity Testing

Butler

2006

Journal of Forensic Sciences

574

350

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Over the past decade, the human identity testing community has settled on a set of core short tandem repeat (STR) loci that are widely used for DNA typing applications. A variety of commercial kits enable robust amplification of these core STR loci. A brief history is presented regarding the selection of core autosomal and Y-chromosomal STR markers. The physical location of each STR locus in the human genome is delineated and allele ranges and variants observed in human populations are summarized as are mutation rates observed from parentage testing. Internet resources for additional information on core STR loci are reviewed. Additional topics are also discussed, including potential linkage of STR loci to genetic disease-causing genes, probabilistic predictions of sample ethnicity, and desirable characteristics for additional STR loci that may be added in the future to the current core loci. These core STR loci, which form the basis for DNA databases worldwide, will continue to play an important role in forensic science for many years to come.

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“…Studies aimed at increasing sensitivity through extract concentration and reduced-volume polymerase chain reaction (PCR) have been performed with a measure of success (6,7). With a standard forensic procedure, the limit of detection for a DNA profile is anywhere from 100 pg to 500 pg, dependent upon the amplification multiplex and detection parameters utilized (8)(9)(10)(11).…”

mentioning

confidence: 99%

Simplified Low‐Copy‐Number DNA Analysis by Post‐PCR Purification

Smith

Ballantyne

2007

Journal of Forensic Sciences

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Frequently, evidentiary items contain an insufficient quantity of DNA to obtain complete or even partial DNA profiles using standard forensic gentotyping techniques. Such low-copy-number (LCN) samples are usually subjected to increased amplification cylces to obtain genetic data. In this study, a 28-cycle polymerase chain reaction (PCR) was used to evaluate various methods of post-PCR purification for their effects on the sensitivity of fluorophore-based allelic detection subsequent to capillary electrophoretic separation. The amplified product was purified using filtration, silica gel membrane, and enzyme mediated hydrolysis purification techniques and evaluated for their effect on fluorescent allelic signal intensity. A purification method was selected and its effect on fluorescent allelic signal intensity was compared with that of the unpurified PCR product. A method of post-PCR purification is described which increases the sensitivity of standard 28-cycle PCR such that profiles from LCN DNA templates (<100 pg DNA) can be obtained. Full DNA profiles were consistently obtained with as little as 20 pg template DNA without increased cycle number. In mock case type samples with dermal ridge fingerprints, genetic profiles were obtained by amplification with 28 cycles followed by post-PCR purification whereas no profiles were obtained without purification of the PCR product. Allele dropout, increased stutter, and sporadic contamination typical of LCN analysis were observed; however, no contamination was observed in negative amplification controls. Post-PCR purification of the PCR product can increase the sensitivity of capillary electrophoresis to such an extent that DNA profiles can be obtained from <100 pg of DNA using 28-cycle amplification.

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Characterization and Validation Studies of Powerplex™ 2.1, A Nine-locus Short Tandem Repeat (Str) Multiplex System and Penta D Monoplex

Cited by 15 publications

References 25 publications

Validation of the multiplex kit genRES MPX-2 for forensic casework analysis

Validation of the multiplex kit genRES MPX-2 for forensic casework analysis

Genetics and Genomics of Core Short Tandem Repeat Loci Used in Human Identity Testing

Simplified Low‐Copy‐Number DNA Analysis by Post‐PCR Purification

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