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PCR DIG Probe Synthesis Kit (25 Reactions, 50 μL Final Volume)
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Specifications:
| Application | PCR | ||
| Storage Temperature | -15°C to -25°C | ||
| Product Type | PCR Reagent | Forms | Kit with Various components |
| Product Brand | ROCHE | ||
| Product Grade | Molecular Biology | ||
For Efficient, High-Sensitivity DIG Labeling of DNA Probes by PCR
The PCR DIG Probe Synthesis Kit provides a reliable and convenient method for the synthesis of digoxigenin (DIG)-labeled DNA probes via polymerase chain reaction (PCR). Incorporating alkali-labile DIG-11-deoxyuridine triphosphate (DIG-11-dUTP) into the DNA backbone, this kit enables robust labeling for sensitive detection in Southern, Northern, and in situ hybridization assays.
Powered by the Expand High Fidelity DNA polymerase mix, the kit enables high-yield labeling of probe fragments ranging from 40 bp to 5 kb, with applications in gene expression analysis, rare mRNA detection, and low-copy gene targeting.
Key Features and Benefits
High Sensitivity DIG Labeling
- Incorporates alkali-labile DIG-11-dUTP at a 1:2 ratio with dTTP.
- Provides enhanced hybridization signals for detection of low-abundance targets.
Robust Amplification with High Fidelity Polymerase
- Includes Expand High Fidelity DNA polymerase mix with proofreading activity.
- Efficiently amplifies and labels from as little as 10 pg plasmid DNA or 10 ng genomic DNA.
Versatile Applications
- Suitable for Southern blots, Northern blots, and in situ hybridization.
- Produces sufficient labeled probe from a single 50 µL reaction to support up to 20 mL of hybridization solution.
Convenient and Flexible Format
- Supplied as a complete 6-component kit.
- Supports adjustable nucleotide concentrations to suit different probe lengths.
Applications
- Low-copy gene detection in genomic Southern blotting
- Rare mRNA expression analysis in Northern blotting
- DNA labeling for in situ hybridization (FISH)
- Probe generation for membrane hybridization assays
- Multiple stripping and reprobing of membranes
Technical Specifications
| Property | Details |
|---|---|
| Product Name | PCR DIG Probe Synthesis Kit |
| SKU / Catalog Number | 11636090910 |
| Reactions per Kit | 25 reactions (50 µL each) |
| Labeling Method | DIG-dUTP incorporation by PCR |
| Template Input Range | 10 pg (plasmid DNA) to 10 ng (genomic DNA) |
| Probe Length Range | 40 bp to 5 kb |
| DIG-dUTP:dTTP Ratio | 1:2 |
| Storage Temperature | –15 to –30 °C (freezer) |
| Shipping Conditions | Room temperature |
| Shelf Life | >1 year (DIG-labeled probe stability) |
| Detection Compatibility | Chemiluminescent, colorimetric, and fluorescent systems |
Kit Components
| Component | Description |
|---|---|
| Enzyme Mix | Expand High Fidelity DNA Polymerase, 3.5 U/μL |
| PCR DIG Probe Synthesis Mix | dATP, dCTP, dGTP (2 mM each), DIG-11-dUTP |
| PCR Buffer with MgCl₂ | 10× concentrated |
| dNTP Stock Solution | dATP, dCTP, dGTP, dTTP (2 mM each), pH 7.0, 10× concentrated |
| Control Template | Plasmid DNA (contains human tPA cDNA), 20 pg/μL |
| Control PCR Primer Mix | Primers 1 and 2, 2 μM each, 50 pmol each |
Quality Assurance
- Each lot is function-tested in PCR and validated by Southern blot hybridization using chemiluminescent detection.
- Control reaction produces a 442 bp DIG-labeled PCR product distinguishable from unlabeled DNA.
The PCR DIG Probe Synthesis Kit offers a sensitive, user-friendly solution for generating DIG-labeled DNA probes suitable for a wide range of hybridization-based applications. With high-performance polymerase, optimized reagents, and validated controls, it ensures consistent results in molecular diagnostics research, transcriptomics, and gene expression profiling.