The Quantifiler Duo DNA Quantification Kit enables forensic laboratories to simultaneously obtain a quantitative and qualitative assessment of total human and human male DNA in a single, highly sensitive real-time PCR reaction. This guides selection of the optimal STR chemistry (autosomal, Y-STR or miniSTR) and streamlines the workflow while increasing downstream analysis success rates.
• Highly Accurate: determines the ratio of human male to female DNA, even with minute male and excess female DNA (1:1000 or greater).
• Highly Informative:
•Guides selection of the optimal AmpFLSTR PCR Amplification Kit (autosomal, Y-STR or miniSTR),
•Improves prediction of STR amplification performance and estimation of DNA quantity in degraded samples,
•Detects the presence of PCR inhibitors (suggesting additional purification of extract and⁄or MiniFiler kit analysis may be necessary).
• Highly Sensitive: 6 pg⁄µl limit of detection.
• Highly Specific: for human and higher primate DNA.
• Highly Efficient: dual quantification system minimizes sample consumption and streamlines workflow.
• Highly Reliable: uses the gold standard TaqMan real-time PCR technology already widely adopted in forensic laboratories.
• Highly Integrated: optimized and validated as part of a complete DNA testing system with the Applied Biosystems 7500 Real-Time PCR System and AmpFLSTR PCR Amplification Kits to provide the highest quality STR profiling results.
A Highly Informative Tool
The Quantifiler Duo kit has been optimized and validated as part of a complete DNA testing system with the AmpFLSTR PCR Amplification Kits.
This enables it to function as a critical component of an improved and integrated workflow for more effective processing of forensic casework samples. The workflow diagram in Figure 1 illustrates how the kit functions as a “sample assessment system" which can efficiently detect the presence of inhibitors, determine the ratio of male to female DNA, and guide selection of the optimal AmpFLSTR PCR Amplification Kit (autosomal, Y-STR or miniSTR) to maximize recovery of interpretable STR profiles.
This kit uses increased amplicon sizes for improved prediction of STR amplification performance and estimation of DNA quantity in degraded samples. The amplicon sizes are similar for all three targets to avoid preferential PCR. The kit uses single copy targets for superior accuracy and consistency in quantification results (multi-copy targets are affected by variability between individuals and decrease correlation with the single copy targets in STR kits).
Comparison to Quantifiler Human and Y Kits
There are many specific differences with the current Quantifiler kits including:
• The Quantifiler Duo kit uses an alternative human DNA target (RPPH1) and increased amplicon sizes to improve prediction of STR amplification performance and estimation of DNA quantity in degraded samples.
• The Quantifiler Duo DNA Standard is pooled human male genomic DNA (rather than cell-line DNA) for improved stability and correlation with forensic samples.
• The Quantifiler Duo PCR Reaction Mix has been optimized for expanded multiplex performance.
• The Quantifiler Duo kit has been designed to guide selection of appropriate AmpFLSTR Kit chemistry and maximize recovery of interpretable STR profiles.