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| Product Name | Loop-Mediated Isothermal Amplification (LAMP) Master Mix |
| Catalog No. | NATR-HMM-0129 |
| Description | A ready-to-use 2× master mix containing Bst DNA polymerase (large fragment) with strong strand-displacement activity, dNTPs, and optimized reaction buffer for isothermal nucleic acid amplification using the LAMP method. The formulation provides robust amplification at a constant temperature, eliminating the need for thermal cycling instrumentation. |
| Intended Use | Rapid, isothermal detection of specific DNA and RNA sequences (with separate reverse transcriptase addition) for point-of-care diagnostics, field-deployable pathogen detection, food safety testing, and molecular biology research applications requiring simplified amplification equipment. |
| Principle / Technology | Bst DNA polymerase large fragment catalyzes DNA synthesis with strand displacement activity at 60–65°C, enabling isothermal amplification without thermal denaturation. The LAMP technique uses four to six primers recognizing six to eight distinct regions on the target sequence, creating a loop-mediated amplification cascade that generates large amounts of DNA with a characteristic stem-loop structure in 15–60 minutes. |
| Detection Method | Real-time fluorescence monitoring using intercalating dyes (SYBR Green I, SYTO 9) or pH-sensitive colorimetric dyes; endpoint detection by turbidity (magnesium pyrophosphate precipitate), color change (phenol red or hydroxynaphthol blue), or agarose gel electrophoresis (characteristic ladder pattern) |
| Sample Type | DNA extracts from clinical specimens (blood, saliva, urine, swabs), food samples, environmental samples, plant tissues, and cultured microorganisms; RNA targets with addition of thermostable reverse transcriptase (RT-LAMP) |
| Performance Range / Specifications | Amplification temperature: 60–65°C (optimally 63°C); reaction time: 15–60 minutes; amplicon detection limit: <10 copies of target DNA per 25 μL reaction; DNA yield: up to 10 μg per 25 μL reaction; detection can be performed visually, by fluorescence, or by turbidity measurement |
| Sensitivity / LOD | Detects as few as 1–10 copies of target nucleic acid per reaction within 30–45 minutes; >100-fold dynamic range for semi-quantitative applications; analytical sensitivity comparable to or exceeding conventional PCR |
| Specificity | High specificity conferred by 4–6 primers targeting multiple recognition sites simultaneously; negligible cross-reactivity with non-target organisms in validated assays; false-positive rate <1% with proper primer design and reagent handling |
| Reaction Conditions / Protocol | Prepare 25 μL reaction: 12.5 μL 2× master mix, LAMP primer set (inner/outer/loop primers at optimized ratios), template DNA, nuclease-free water to volume; incubate at 63°C for 30–60 minutes in heat block, water bath, or real-time instrument; optional heat inactivation at 80°C for 5 minutes; observe color change or measure fluorescence for result interpretation |
| Components / Formulation | Bst DNA polymerase large fragment (from Geobacillus stearothermophilus, recombinant), dNTPs (dATP, dCTP, dGTP, dTTP), Tris-HCl, KCl, (NH4)2SO4, MgSO4, Triton X-100, proprietary enhancer blend, pH-sensitive dye (phenol red or neutral red) in colorimetric formulations, and thermostable reverse transcriptase in RT-LAMP formulations |
| Storage Conditions | –20°C for long-term storage; stable at 2–8°C for up to 1 month; room temperature for up to 24 hours during use; minimize freeze-thaw cycles; protected from light for colorimetric dye-containing formulations |
| Shelf Life | 18 months at –20°C from date of manufacture |
| Package Specifications | 120 reactions (based on 25 μL), 480 reactions, and custom bulk formats for high-throughput applications |
| Product Form | Liquid 2× master mix, may include colorimetric dye version for visual detection |
| Quality Control | Each lot tested: amplification of 10 copies of synthetic DNA target within 30 minutes at 63°C; no amplification in no-template control (60-minute incubation); RT-LAMP variant tested for RNA target with MS2 phage RNA; visual color change (pink → yellow for phenol red) clear and unambiguous; inter-lot consistency in time-to-positive <5% CV |
| Key Features | No thermal cycler required — operates isothermally at 63°C; visual colorimetric readout option for instrument-free detection; amplification times as short as 10–15 minutes; high tolerance to biological inhibitors in crude samples; compatible with lyophilization for ambient temperature storage and transport |
For research use only, not for clinical use.
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