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| Product Name | RNA Gel Loading Dye, 2×, Denaturing, with Ethidium Bromide |
| Catalog No. | NATR-HMM-0151 |
| Description | 2× denaturing RNA gel loading dye containing formamide, formaldehyde, and ethidium bromide for visualization of RNA on denaturing agarose gels. Formamide and formaldehyde maintain RNA in a denatured, single-stranded state to ensure migration proportional to molecular weight rather than secondary structure. Pre-mixed ethidium bromide eliminates post-electrophoresis staining, enabling direct RNA visualization under UV light after electrophoresis. |
| Intended Use | Loading of RNA samples onto formaldehyde-agarose denaturing gels for Northern blotting and RNA integrity assessment; denaturing RNA electrophoresis for size verification; RNA quality control prior to cDNA synthesis and NGS library preparation. |
| Principle / Technology | Formamide (50% v/v) and formaldehyde (6.5% v/v) denature RNA secondary structure; EDTA chelates Mg²⁺ which catalyzes RNA hydrolysis at elevated temperatures; ethidium bromide intercalates into RNA enabling direct UV visualization (no post-stain required); bromophenol blue tracks electrophoresis progress (~500 nt migration) |
| Detection Method | Mix RNA sample (1-5 µg in ≤10 µL) with equal volume 2× loading dye; heat at 65 °C for 10 minutes to denature; chill on ice 2 minutes; load onto formaldehyde-agarose gel (1-1.5% agarose, 6.5% formaldehyde in MOPS buffer); run at 5-7 V/cm until bromophenol blue reaches 2/3 gel length |
| Sample Type | Total RNA (1-5 µg per lane) or mRNA (0.5-2 µg); RNA must be in nuclease-free water or TE buffer; samples with salts (>50 mM) or ethanol may cause loading problems |
| Sensitivity / LOD | Visualizes 0.5-5 µg total RNA per lane under UV transillumination (28S and 18S rRNA bands visible); ethidium bromide detection limit ~10 ng RNA per band |
| Specificity | Formamide + formaldehyde + heat denaturation ensures complete RNA denaturation; single-stranded RNA migrates proportionally to molecular weight; ethidium bromide stains ssRNA (lower efficiency than dsDNA but sufficient for typical loads) |
| Reaction Conditions / Protocol | Mix 1:1 with RNA sample; heat 65 °C 10 min; chill on ice 2 min; load immediately — extended time on ice may allow partial renaturation |
| Components / Formulation | Formamide (50% v/v), formaldehyde (6.5% v/v, from 37% stock), ethidium bromide (10 µg/mL), bromophenol blue (0.02% w/v), MOPS buffer (1×: 20 mM MOPS, 5 mM sodium acetate, 1 mM EDTA, pH 7.0), glycerol (10% v/v) |
| Storage Conditions | -20 °C; protect from light; formamide/formaldehyde mixture is moisture-sensitive — keep tightly sealed; ethidium bromide is a mutagen — handle with gloves |
| Shelf Life | 12 months at -20 °C; use within 3 months after first thaw |
| Package Specifications | 1 mL, 5 × 1 mL aliquots in amber tubes |
| Product Form | Dark blue/purple liquid, ready-to-use; aliquot format to minimize freeze-thaw and contamination |
| Key Features | Complete denaturing dye — formamide + formaldehyde + heat; pre-mixed ethidium bromide for direct visualization; bromophenol blue tracking dye; MOPS buffer compatibility for Northern blotting; 28S:18S rRNA ratio (2:1) verifies RNA integrity; amber tubes for light protection; ready-to-use |
| Purity | Formamide deionized, conductivity <10 µS/cm; formaldehyde methanol-free (<1% methanol); MOPS buffer components molecular biology grade; DNase/RNase tested free |
| Concentration | 2× concentrate; use 1:1 dilution with RNA sample |
| Activity / Unit Definition | Polymerase activity defined as nmol dNTP incorporated per 30 min per unit; other activities per product specification |
| Molecular Weight | As specified per DNA markers or protein components |
| Source / Origin | Recombinant enzymes expressed in E. coli; synthetic oligonucleotides; ultrapure reagents |
| pH Range / Optimal pH | pH 7.0 ± 0.1 (MOPS buffer component) |
| Shipping Conditions | Ambient temperature; ethidium bromide is a mutagen — appropriate labeling and packaging per IATA regulations |
| Expiration Date / Stability | 12 months at -20 °C; 3 months after first thaw; formamide oxidizes over time to formic acid — discard if pH drops >0.5 units (measured by 10-fold dilution in water); discard if ethidium bromide fluorescence decreases significantly |
| Regulatory / Compliance | For laboratory and research use only; RUO; ethidium bromide is a potent mutagen — handle with nitrile gloves and dispose as hazardous chemical waste; formamide is a reproductive toxin; formaldehyde is toxic and carcinogenic — use in chemical fume hood; ISO 9001 |
| Compatibility | Compatible with 1-1.5% formaldehyde-agarose gels in 1× MOPS buffer; not compatible with TAE or TBE buffer systems (formaldehyde requires MOPS buffer for effective denaturation); gel must contain 6.5% formaldehyde; running buffer must be 1× MOPS with 6.5% formaldehyde; ethidium bromide can be omitted for downstream Northern blotting (probe detection) — use ethidium-free version if post-electrophoresis hybridization is planned |
| Recommended Buffer System | Tris-HCl, KCl, MgCl₂-based reaction buffer; TE for nucleic acid storage |
| Application Notes / Precautions | Use dedicated PCR workspace; aliquot enzymes to avoid contamination; monitor no-template controls |
| Batch-to-Batch Consistency | Enzyme activity within ±15% of reference lot; PCR performance verified with control template |
For research use only, not for clinical use.
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