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| Product Name | Real-Time PCR Sample Diluent, Low-EDTA TE Buffer, Nuclease-Free |
| Catalog No. | ATRD-0025 |
| Description | Nuclease-free low-EDTA TE buffer optimized for dilution of DNA and RNA templates prior to real-time quantitative PCR (qPCR) and digital PCR (dPCR). The reduced EDTA concentration (0.1 mM) prevents magnesium chelation interference with DNA polymerase activity while still providing sufficient nuclease inhibition. Formulated with molecular biology grade Tris and EDTA, processed through 0.1 µm filtration and certified free of detectable DNase, RNase, and human genomic DNA contamination. Ideal for serial dilutions of nucleic acid standards in qPCR calibration curves. |
| Intended Use | Dilution of nucleic acid templates (DNA, cDNA, RNA) for quantitative real-time PCR, digital PCR, and standard curve preparation in gene expression analysis, genotyping, and pathogen detection workflows. |
| Principle / Technology | Tris buffer maintains pH 8.0 for nucleic acid stability; low EDTA (0.1 mM) chelates trace divalent cations that activate nucleases while preserving free Mg2+ for polymerase activity; nuclease-free certification ensures template integrity. |
| Detection Method | Prepare serial dilutions of nucleic acid template in diluent; use immediately or store at -20 °C; include diluent-only wells as no-template controls (NTC) in each PCR run. |
| Sample Type | Purified genomic DNA, plasmid DNA, cDNA, RT-PCR products, synthetic oligonucleotide templates. |
| Performance Range / Specifications | Maintains nucleic acid integrity for >24 hours at 4 °C and >12 months at -20 °C; NTC wells consistently negative through 45 amplification cycles with 5 µL template. |
| Sensitivity / LOD | Free Mg2+ concentration of 0.1 mM EDTA is below detection threshold for PCR interference (typically requires >0.5 mM EDTA in reaction). |
| Specificity | Formulated specifically for qPCR compatibility; low EDTA minimizes Mg2+ sequestration that could reduce polymerase efficiency. |
| Reaction Conditions / Protocol | Dilute template to desired concentration; use 2-5 µL diluted template per 20 µL PCR reaction. |
| Components / Formulation | Tris-HCl 10 mM, EDTA 0.1 mM, pH 8.0, nuclease-free water (18.2 MΩ·cm). |
| Storage Conditions | Store at room temperature (15-25 °C) or 2-8 °C; aliquot and store at -20 °C for long-term storage. |
| Shelf Life | 24 months from date of manufacture in unopened container. |
| Package Specifications | 50 mL, 100 mL, 500 mL, 1 L nuclease-free bottles. |
| Product Form | Clear, colorless liquid; sterile-filtered 0.1 µm. |
| Quality Control | Each lot tested for DNase activity (DNaseAlert), RNase activity (RNaseAlert), human genomic DNA contamination (Alu qPCR), pH verification, and endotoxin. |
| Key Features | Low EDTA (0.1 mM) for qPCR compatibility; nuclease-free certified; human gDNA-free; 0.1 µm filtered; long shelf life at room temperature; suitable for digital PCR applications. |
| Purity | Molecular biology grade Tris and EDTA; water resistivity 18.2 MΩ·cm at 25 °C; TOC <10 ppb. |
| Concentration | 1× ready-to-use; Tris-HCl 10 mM, EDTA 0.1 mM. |
| Activity / Unit Definition | Nuclease absence verified by lack of fluorescence increase in nuclease substrate assays after 24-hour incubation. |
| Molecular Weight | Tris: 121.14 g/mol; EDTA disodium: 372.24 g/mol. |
| Source / Origin | Synthetic molecular biology grade reagents; Ultrapure water from validated water purification system. |
| pH Range / Optimal pH | pH 8.0 ± 0.1 at 25 °C. |
| Shipping Conditions | Ambient temperature. |
| Expiration Date / Stability | 24 months at room temperature; retest annually for nuclease activity for extended storage. |
| Regulatory / Compliance | For research use only; not for diagnostic procedures. Manufactured in ISO 13485 certified facility. |
| Compatibility | Compatible with all major qPCR master mixes: SYBR Green, TaqMan probe-based, and intercalating dye chemistries. Compatible with digital PCR oil-based partitioning systems. |
| Recommended Buffer System | 10 mM Tris-HCl buffer, pH 8.0. |
| Application Notes / Precautions | Aliquot working volume to minimize freeze-thaw cycles and reduce contamination risk. Always prepare no-template controls using the same diluent batch. For RNA templates, work quickly and keep samples on ice to minimize RNase exposure. Add carrier RNA (5-10 ng/µL) for very low concentration DNA templates to minimize surface adsorption losses. |
| Batch-to-Batch Consistency | pH within ±0.1 units; nuclease testing negative for each lot; endotoxin <0.05 EU/mL. |
For research use only, not for clinical use.
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