The AmpFlSTR Identifiler Plus PCR Amplification Kit and the AmpFlSTR Identifiler Direct PCR Amplification Kit are short tandem repeat (STR) multiplex assays that amplifiy 15 tetranucleotide repeat loci and the amelogenin gender-determining marker in a single PCR amplification. The kits contain the original 13 required loci for the Combined DNA Index System (CODIS) and two additional loci, D2S1338 and D19S433, to ensure compatibility with additional global database requirements.
These kits also maintain primer sequences to enable genotyping concordance for samples previously typed using the Identifiler, Profiler Plus, COfiler, or SGM Plus kits, and facilitate comparison with existing DNA databases. The kits are manufactured in our location in Warrington, UK, a facility that meets the guidelines for ISO 18385 certification and are supported by our comprehensive training, service, and applications teams.
AmpFlSTR Identifiler Plus PCR Amplification Kit
The AmpFlSTR Identifiler Plus PCR Amplification Kit has been developed specifically to address the needs of forensic casework laboratories. It utilizes the same primers as the widely used AmpFlSTR Identifiler PCR Amplification Kit and harnesses next-generation PCR amplification technology to help provide a new level of performance, data quality, and efficiency. This enables forensic analysts to recover more interpretable results from challenging casework samples with increased confidence.
- Helps significantly improve performance on severely inhibited casework samples.
A comparison of the performance of the Identifiler Plus kit and the Identifiler kit on an inhibited casework sample. The Identifiler kit is unable to amplify a blood sample from a leather glove. With its enhanced formulation, the Identifiler Plus kit helps deliver a full profile. For both kits, 1 ng of input DNA and 28 cycles of amplification were used.
- Helps overcome PCR inhibition through the use of a high performance PCR buffer system and optimized thermal cycling parameters.
A study demonstrating the performance of the Identifiler Plus kit in the presence of increasing concentrations of common PCR inhibitors hematin and humic acid. Even at the highest concentrations, the Identifiler Plus kit shows the ability to recover full profiles. 1 ng input DNA and 28 cycles of amplification were used for all replicates.
- Yields higher peak heights from low-level samples and minor contributors in mixtures to facilitate interpretation
An example of a casework sample that yielded no profile when analyzed with the Identifiler kit but yielded a high-quality, interpretable mixture profile with the Identifiler Plus kit. For both kits, 1 ng of input DNA and 28 cycles of amplification were used.
- Enables greater sensitivity compared to the Identifiler kit and provides additional analytical flexibility by utilizing fully-validated 28- and 29-cycle protocols
Illustration of the improved sensitivity of the Identifiler Plus kit. This figure shows the average peak heights (rfu) obtained for four different dilutions of a control DNA sample amplified for 28 cycles using the Identifiler kit (green bars) and Identifiler Plus kit (orange bars). On average, the Identifiler Plus kit peak heights were approximately 1.7 times higher than Identifiler kit peak heights.
- Streamlines workflow by reducing amplification time by one hour, compared to the Identifiler kit.
- Maintains the small amplicon size range of the Identifiler kit (<360 bp) for recovery of information from degraded samples; works in conjunction with the AmpFLSTR MiniFiler PCR Amplification Kit to help maximize recovery from heavily degraded samples.
- Maintains primer sequences for genotyping concordance with samples typed with the Identifiler, Identifiler Direct, and AmpFLSTR SGM Plus kits.
- Utilizes an improved primer manufacturing process to help significantly reduce artifacts, even at increased cycle number.
A comparison of negative control reactions for the Identifiler kit at 28 amplification cycles, the Identifiler Plus kit at 28 amplification cycles and the Identifiler Plus kit at 29 amplification cycles. The improved primer manufacturing process incorporated into the Identifiler Plus kit enables users to obtain significantly cleaner baselines.
AmpFlSTR Identifiler Direct PCR Amplification Kit
The AmpFlSTR Identifiler Direct PCR Amplification Kit has been developed to automate the front end of the single-source sample workflow, streamlining the entire process, while maximizing sampling integrity and reliability. By eliminating the tedious steps involved in DNA extraction and purification, the Identifiler Direct kit facilitates high-throughput processing with a simple, easy-to-automate protocol that requires a less sophisticated and less expensive robotic platform.
- Helps eliminate the need for extraction and purification of blood or buccal samples on FTA paper, enabling users to just "punch and go".
- Accelerates the time to result while maintaining high profile quality
- Reduces the number of pre-amplification processing steps, which minimizes the possibility of contamination or other procedural errors
- Helps facilitate high-throughput processing with a simple, easy-to-automate protocol that requires a less sophisticated and less expensive robotic platform.
Example of a blood sample on FTA card amplified using the Identifiler Direct kit. Heterozygote peak heights average approximately 2000 rfu.
DNA extracted from liquid blood (using Applied Biosystems BloodPrep chemistry on a 6100 platform), with 1 ng amplified using the Identifiler kit compared to the same blood samples immobilized on FTA cards and direct-amplified using the Identifiler Direct kit.
Combined results from four U.S. test site laboratories that tested the Identifiler Direct kit with blood and buccal samples on FTA cards.
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