PCR Amplification for Forensic DNA Profiling
DNA profiling has been used in forensics since 1985 when minisatellites and Southern blotting were the technology of the day. In the early 1990s, short tandem repeats (STRs) were introduced as a new polymorphic DNA marker type and have since become the gold standard for human identification in DNA databases, criminal casework, paternity and kinship analyses, and identification of missing persons. Recent advances in designing large multiplex STR panels have greatly increased the power of discrimination to aid in inclusion and exclusion decisions in human identification applications. Regional requirements for specific markers vary but with ongoing international data sharing efforts, STR PCR amplification kits need maximum commonality in markers to make more cross-border identifications.
PCR amplification products
GlobalFiler STR Kits combine reduced amplification time with maximum data recovery power. The 6-dye, 24-locus technology is designed to deliver superior lab performance. Both the GlobalFiler and GlobalFiler Express kits are approved for use by laboratories generating DNA profiles for inclusion in the US National DNA Index System (NDIS) CODIS database.
Y-STR analysis focuses on short tandem repeats (STRs) found on the Y chromosome, only carried by male individuals. For forensic labs, the most common application of Y-STRs is in criminal cases such as sexual assault, where evidence samples are difficult to resolve with traditional autosomal STR analysis due to low amounts of male DNA present in a high background of female DNA.
Optimized for casework, the AmpFℓSTR™ Identifiler™ Plus PCR Amplification Kit enables forensic analysts to recover more interpretable results from a wider range of DNA evidence samples with increased confidence.
VeriFiler STR Kits offer superior genotyping results from single-source and challenging casework samples using a 25-marker STR multiplex assay that includes two gender determination markers, as well as the highly discriminatory Penta D and Penta E markers.
The NGM Detect PCR Amplification Kit offers superior sensitivity to maximize recovery of data from casework samples. Powered by 6-dye Chemistry, the new NGM Detect kit builds on the well established NGM SElect format, with new primer sequences that enable optimal marker positioning developed specifically for users who require loci based on ENFSI and EDNAP requirements. Together, the two kits offer a powerful combination to gather maximal information from challenging samples.
NGM™ Kits are designed to deliver exceptional data quality and robustness along with powerful discrimination to support European cross-border data sharing initiatives. The NGM™ kit amplifies all 10 SGM Plus™ kit loci together with the 5 additional loci in the expanded European Standard Set.
The AmpFℓSTR™ NGM Select™ Express Kit enables extremely efficient analysis of single-source samples for laboratories wishing to utilize the European Standard Set Loci (ESSL) together with the highly discriminating SE33 locus.
The Quantifiler Trio kit, used in conjunction with the HID Real-Time PCR Analysis Software, redefines the casework sample-processing workflow. Quality index functionality enables a more comprehensive view of sample quality (both degradation and inhibition), while increased sensitivity and robustness correlate with more recently released Applied Biosystems STR chemistries. Screen samples and make critical workflow decisions (for example, autosomal vs. Y-STR) with greater confidence.
Building on the reliability and performance of Applied Biosystems™ thermal cycler instruments, the HID Veriti™ Thermal Cycler features a 96-well (0.2 mL) format with six independent temperature zones to provide precise temperature control and rapid temperature changes for conducting PCR.
The ProFlex PCR System combines the reliability and performance of Applied Biosystems™ thermal cyclers with the flexible configuration and control features that fit how you work today. Dual block configurations allow you to maximize your throughput.
Flexibility of 6-dye Chemistries in recovering maximum information
Dr. Antonio Alonso, a DNA expert at the National Institute of Toxicology and Forensic Sciences, Madrid (Spain), discusses the need for forensic laboratories to make the transition to 6-dye Chemistry technology in order to gain workflow efficiency and reduce operating costs. He highlights the value of Y-STR analysis in resolving sexual assault cases and provides the example of an 18-year-old rape/murder cold case in Spain that was solved using Applied Biosystems™ Yfiler™ Plus chemistry.
Value of Y-STRs in resolving mixture samples
Y-STR analysis can increase the success rate of investigations into sexual assaults, particularly in cases where only non-sperm traces may be available such as saliva, skin, finger nails, contact & touch traces. Such mixture samples contain small amounts of male DNA in a high background of female DNA and autosomal testing often does not yield a complete profile. Dr. Franz Neuhuber, Head of Forensic Molecular Biology Department at University of Salzburg, Austria shares details of an attempted rape case that prompted the lab to adopt Y-STR analysis.
For Research Forensic or Paternity Use Only. Not for use in diagnostic procedures. For licensing and limited use restrictions visit thermofisher.com/HIDlicensing