Quantitation, Normalization and PCR Setup

Following extraction, forensic DNA samples are usually quantified to determine the amount of human DNA present, then the amount of input template DNA for the STR amplification is normalized based on these quantitation values. With increased automation of DNA extraction there is a concomitant need to increase automation of these downstream processes to prevent a bottleneck from forming in the workflow at these stages.

Products for Human DNA Quantitation

The Plexor® HY System
Promega is currently evaluating automation of the Plexor® HY System, a real-time PCR method that simultaneously quantifies total human DNA and male DNA. This system uses the Plexor® technology, which results in decreasing fluorescence as the amplification progresses, and provides consistent DNA detection down to 6.4pg. Contact genetic@promega.com for information about automated Plexor® HY setup methods.

Learn more about the Plexor® HY System

Products for STR Amplification

PowerPlex® 16 System
The PowerPlex® 16 System allows amplification of all 13 CODIS loci, in addition to amelogenin and the highly polymorphic Penta D and Penta E loci in a single amplification. The amplified products can be resolved using an ABI PRISM® 310, 3100 or 3100-Avant Genetic Analyzer, ABI PRISM® 377 DNA Sequencer or Applied Biosystems 3130 or 3130xl Genetic Analyzer. Data can be subsequently analyzed using the GeneScan®/GenoTyper® or GeneMapper® ID software or with the FSS-i3™ Expert Systems Software.

Learn more about the PowerPlex® 16 System

PowerPlex® Y System
The PowerPlex® Y System allows amplification of 12 Y-STR loci, including all loci in the "European Minimal Haplotype" and SWGDAM-recommended Y-STR panel in a single amplification. The amplified products can be resolved using an ABI PRISM® 310, 3100 or 3100-Avant Genetic Analyzer, ABI PRISM® 377 DNA Sequencer or Applied Biosystems 3130 or 3130xl Genetic Analyzer. Data can be subsequently analyzed using GeneScan®/GenoTyper® or GeneMapper® ID software or with the FSS-i3™ Expert Systems Software.

Learn more about the PowerPlex® Y System

PowerPlex® 16 BIO System
The PowerPlex® 16 BIO System allows amplification of all 13 CODIS loci, in addition to amelogenin and the highly polymorphic Penta D and Penta E loci in a single amplification. Unlike the PowerPlex® 16 System, the PowerPlex® 16 BIO System is designed specifically for use on the Hitachi FMBIO® II Fluorescence Imaging System with StaR Call™ software.

Learn more about the PowerPlex® 16 BIO System

PowerPlex® ES System
The PowerPlex® ES System allows amplification of all 7 ENFSI loci, in addition to amelogenin and the highly polymorphic SE33 locus in a single amplification. The amplified products can be resolved using an ABI PRISM® 310, 3100 or 3100-Avant Genetic Analyzer, ABI PRISM® 377 DNA Sequencer or Applied Biosystems 3130 or 3130xl Genetic Analyzer. Data can be subsequently analyzed using GeneScan®/GenoTyper® or GeneMapper® ID software or with the FSS-i3™ Expert Systems Software.

Learn more about the PowerPlex® ES System

Promega-Supported Platforms for DNA Quantitation, Normalization and PCR Setup

Promega has worked in conjunction with Beckman Coulter to develop an automated solution for DNA quantitation, normalization and PCR setup on the Beckman Coulter Biomek® 2000 Laboratory Automation Workstation.

NonPromega-Supported Instrument Platforms for DNA Quantitation, Normalization and PCR Setup

Customers and other instrument suppliers have developed their own methods for human DNA quantitation, normalization and PCR setup on other instruments. Though these methods have not been validated by Promega scientists, they might be of interest to those interested in automation. PerkinElmer has developed a method for DNA quantitation, subsequent normalization and PCR setup on their JANUS® Automated Workstation. This method is installed and fully supported by PerkinElmer.

For more details please see JANUS® Forensic Workstation (pdf: 253KB) and the PerkinElmer web site.