PerfeCTa Multiplex qPCR ToughMix
Features & Benefits
- 1-tube SuperMix minimizes pipetting steps, simplifying reaction assembly and improving accuracy
- 5x concentrated reagent provides more flexibility with dilute DNA samples sensitivity
- ToughMix reagent technology neutralizes a broad spectrum of PCR inhibitors common in plant and animal tissues, environmental sources, clinical specimens and complex food matrices
- Superior assay sensitivity and specificity with ultrapure AccuStart II enzyme technology – maximum-yielding Taq DNA polymerase mutant controlled by stringent, multi-epitope antibody hotstart
- Supports efficient vortex mixing with proprietary anti-foaming additives
PerfeCTa Multiplex qPCR ToughMix is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.
Suppression of low copy amplicons by high copy reference targets during multiplex co-amplification skews the apparent representation and quantification of low copy target sequences. PerfeCTa MultiPlex qPCR ToughMix transcends these limitations by enabling sensitive, broad linear dynamic detection range with co-amplification of four abundant (106) targets. PerfeCTa MultiPlex qPCR ToughMix enables multiplex qPCR assay performance comparable to single-plex qPCR probe assays without the need to rigorously titrate primer concentration. PerfeCTa MultiPlex qPCR ToughMix is a 5x concentrated, ready to-use reaction cocktail for real-time quantitative PCR (qPCR) with ToughMix reagent technology that neutralizes a broad spectrum of PCR inhibitors that compromise assay performance with crude extracts, clinical specimens, plant, soil environmental or complex food matrices. A key component of the kit is an ultrapure, highly-processive thermostable DNA polymerase that is free of detectable E. coli DNA. PPerfeCTa MultiPlex qPCR ToughMix is ideal for demanding qPCR applications such as bacterial pathogen detection where residual host DNA in typical recombinant enzyme preparations can limit assay sensitivity and obscure detection of low copy samples.
A Multiplexed Real-time PCR Assay to Detect SCID and SMN1 Homozygous Exon 7 Deletion, and A Droplet Digital PCR Assay to Assess SMN2 Copy Numbers, in Newborn screening for Spinal Muscular Atrophy
Single-tube, 5X concentrated reagent containing:
- Reaction buffer with optimized concentrations of molecular-grade MgCl2, dATP, dCTP, dGTP, dTTP.
- AccuStart II Taq DNA Polymerase.
- Proprietary enzyme stabilizers and performance-enhancing additives.
- Titrated reference dye (if applicable).
Storage & HandlingStore components in a constant temperature freezer at -25°C to -15°C protected from light upon receipt. Repeated freezing and thawing does not impair product performance. For lot specific expiry date, refer to package label, Certificate of Analysis or Product Specification Form.
- Applied Biosystems 5700
- Applied Biosystems 7000
- Applied Biosystems 7300
- Applied Biosystems 7700
- Applied Biosystems 7900
- Applied Biosystems 7900HT
- Applied Biosystems 7900 HT Fast
- Applied Biosystems StepOne™
- Applied Biosystems StepOnePlus™
- Applied Biosystems 7500
- Applied Biosystems 7500 Fast
- Stratagene Mx3000P®
- Stratagene Mx3005P™
- Stratagene Mx4000™
- Applied Biosystems ViiA 7
- Applied Biosystems QuantStudio™
- Agilent AriaMx
- Douglas Scientific IntelliQube®
Bio-Rad iCycler iQ systems
- Quantabio Q
- BioRad CFX
- Roche LightCycler 480
- QIAGEN Rotor-Gene Q
- BioRad iCycler iQ™
- BioRad MyiQ™
- BioRad iQ™5
Related ResourcesProduct FlyersProduct ManualsPerfeCTa MultiPlex qPCR ToughMix PerfeCTa MultiPlex qPCR ToughMix, ROXPerfeCTa MultiPlex qPCR ToughMix, Low ROXTechnical NotesPublicationsDetection and Quantification of Acute Myeloid Leukemia-Associated Fusion TranscriptsJonathan Schumacher, Acute Myeloid Leukemia - 2017AbstractReal-time quantitative reverse transcription polymerase chain reaction (RT-qPCR)-based detection of abnormal fusion transcripts is an important strategy for the diagnosis and monitoring of patients with acute myeloid leukemia (AML) with t(8;21)(q22;q22); RUNX1-RUNX1T1, inv(16)(p13.1;q22); CBFB-MYH11 or t(15;17)(q22;q12); PML-RARA. In RT-qPCR assays, patient-derived cDNA is subjected to amplification using PCR primers directed against the fusion transcript of interest as well as a reference gene for normalization. Quantification is typically performed by constructing standard curves for each PCR run using a series of plasmid standards of known concentration that harbor the same fusion transcript or the same reference gene of interest. Fusion transcripts and reference gene copy numbers are then calculated in patient samples using these standard curves. The process of constructing standard curves is laborious and consumes additional reagents. In this chapter, we give the method details for a multiplex RT-qPCR strategy to detect and quantify the acute myeloid leukemia (AML)-associated fusion transcripts PML-RARA in patients with t(15;17) without the need for standard curves. This general method can also be applied to other AML-associated fusion transcripts such as CBFB-MYH11 and RUNX1-RUNX1T1.Crystal digital droplet PCR for detection and quantification of circulating EGFR sensitizing and resistance mutations in advanced non-small cell lung cancerCécile Jovelet, PLOS ONE - 2017AbstractOver the past years, targeted therapies using tyrosine kinase inhibitors (TKI) have led to an increase in progression-free survival and response rate for a subgroup of non-small cell lung cancer (NSCLC) patients harbouring specific gene abnormalities compared with chemotherapy. However long-lasting tumor regression is rarely achieved, due to the development of resistant tumoral subclones, which requires alternative therapeutic approaches. Molecular profile at progressive disease is a challenge for making adaptive treatment decisions. The aim of this study was to monitor EGFR-mutant tumors over time based on the quantity of mutant DNA circulating in plasma (ctDNA), comparing two different methods, Crystal™ Digital™ PCR and Massive Parallel Sequencing (MPS). In plasma circulating cell free DNA (cfDNA) of 61 advanced NSCLC patients we found an overall correlation of 78% between mutated allelic fraction measured by Crystal Digital PCR and MPS. 7 additional samples with sensitizing mutations and 4 additional samples with the resistance mutation were detected with Crystal Digital PCR, but not with MPS. Monitoring levels of both mutation types over time showed a correlation between levels and trends of mutated ctDNA detected and clinical assessment of disease for the 6 patients tested. In conclusion, Crystal Digital PCR exhibited good performance for monitoring mutational status in plasma cfDNA, and also appeared as better suited to the detection of known mutations than MPS in terms of features such as time to results.Prevalence and associated risk factors of