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| Product Name | TaqMan Probe Custom Synthesis Service |
| Catalog No. | PRIPRO-0014 |
| Description | A comprehensive custom TaqMan probe synthesis service for quantitative real-time PCR assay development. This service covers the full workflow from probe sequence design assistance through synthesis, dual-labeling, HPLC purification, and quality control. Users can select from a wide range of 5′ reporter fluorophores (FAM, HEX, TET, JOE, ROX, TAMRA, CY3, CY5, Texas Red, and others) paired with 3′ quenchers (BHQ1, BHQ2, BHQ3, TAMRA, DABCYL). The service includes expert design review to ensure probe Tm, GC content, secondary structure, and specificity meet assay requirements. Each probe undergoes dual-HPLC purification, MALDI-TOF mass spectrometry, and functional validation of quenching efficiency. A detailed certificate of analysis is provided with each batch. This service supports applications from single-gene expression assays to high-order multiplex panels. |
| Intended Use | Designed for researchers developing custom qPCR assays for gene expression quantification, pathogen detection, SNP/mutation genotyping, copy number variation analysis, microRNA quantification, and multiplex diagnostic panel development. |
| Principle / Technology | TaqMan probe chemistry: dual-labeled oligonucleotide with 5′ reporter and 3′ quencher; fluorescence generated by Taq polymerase 5′-3′ exonuclease cleavage during PCR extension |
| Detection Method | Real-time fluorescence detection; probe performance validated by standard curve qPCR with synthetic template to confirm efficiency, linearity, and sensitivity |
| Sample Type | Custom sequence provided by user or designed collaboratively; applicable to genomic DNA, cDNA, plasmid DNA, or synthetic templates |
| Performance Range / Specifications | Probe length 15–35 bases; Tm 65–70 °C (5–10 °C above primer Tm); GC 30–80%; no 5′ G; avoid runs of >4 identical bases; qPCR efficiency 90–110% when paired with optimized primers |
| Sensitivity / LOD | Detection limit ≤10 copies of target per reaction with properly designed probe-primer set; dynamic range exceeding 6 orders of magnitude |
| Specificity | Sequence-specific detection; no signal from non-target templates; BLAST-verified probe sequence uniqueness included in design review |
| Reaction Conditions / Protocol | Final probe concentration 100–250 nM in 20–50 µL qPCR reaction. Recommend two-step cycling: 95 °C 15 s, 60 °C 30–60 s with fluorescence acquisition. Optimization service available. |
| Components / Formulation | Custom-synthesized dual-labeled HPLC-purified probe; design report with in silico analysis; certificate of analysis; optional primer pair synthesis available |
| Storage Conditions | –20 °C protected from light; dry or in TE buffer aliquots |
| Shelf Life | 12 months at –20 °C dry (dark); 6 months at –20 °C in solution (dark) |
| Package Specifications | Light-protective vial; scale options from 5 nmol to 1 µmol; custom packaging upon request |
| Product Form | Lyophilized dual-labeled oligonucleotide; color varies by fluorophore choice |
| Quality Control | HPLC ≥95% dual-labeled purity; MALDI-TOF MS mass confirmation; quenching efficiency assay; optional functional qPCR validation; endotoxin testing available. |
| Key Features | Full design support with in silico specificity check; wide selection of fluorophore-quencher combinations; HPLC purified to >95%; MS verified; optional functional validation; rapid turnaround within 5–7 business days for standard probes. |
| Purity | ≥95% dual-labeled by HPLC; free dye <3%; single-labeled oligo <2% |
| Concentration | Resuspend to 100 µM stock; yield quantified by A260 with dye absorbance correction; scale-dependent yield reported on COA |
| Activity / Unit Definition | Quenching efficiency >95%; end-point fluorescence increase >10-fold upon nuclease digestion; functional qPCR efficiency 90–110% when optimized |
| Molecular Weight | Calculated per sequence plus dye/quencher modifications; verified by MS |
| Source / Origin | Synthetic; all raw materials from qualified chemical suppliers |
| pH Range / Optimal pH | qPCR optimal pH 8.0–8.5; probe chemically stable pH 6.0–9.0 |
| Shipping Conditions | Ambient temperature; light-protected; expedited shipping available |
| Expiration Date / Stability | 12 months at –20 °C dry, protected from light; 6 months at –20 °C in TE buffer; stability study data available for specific modifications |
| Regulatory / Compliance | ISO 9001 certified facility; for research use only; custom ISO 13485 documentation available for IVD development projects |
| Compatibility | Compatible with all major qPCR instruments including ABI, Bio-Rad, Roche, Qiagen, Agilent platforms; all commercially available qPCR master mixes |
| Recommended Buffer System | Design review includes buffer compatibility check; standard TE or nuclease-free water for resuspension |
| Application Notes / Precautions | Submit probe target sequence with amplicon context for design review. For multiplex assays, indicate all fluorophores to avoid spectral overlap. Contact technical support for sequences with challenging GC content or secondary structure. |
| Batch-to-Batch Consistency | HPLC purity maintained ≥95% per batch; quenching efficiency within ±3%; mass accuracy ±0.05% across all lots |
For research use only, not for clinical use.
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