DNA/RNA Stain | MIDORI Green Direct

1. Product Overview - MIDORI Green Direct – Direct DNA/RNA Staining with Integrated Loading Dye

MIDORI Green Direct is a fluorescent nucleic acid stain developed as an alternative to ethidium bromide (EtBr) for DNA and RNA detection by agarose gel electrophoresis. Unlike stains that must be incorporated into the agarose gel or running buffer, MIDORI Green Direct is added directly to the sample before loading. The formulation already contains loading dye, simplifying sample preparation and reducing the number of handling steps in the laboratory.

The stain can be used to visualize double-stranded DNA (dsDNA), single-stranded DNA (ssDNA), and RNA. It is optimized for blue LED and blue/green LED illumination and can also be used with conventional UV transilluminators, although UV is not the manufacturer's preferred option.

Key Benefits

• Direct sample staining: Add the stain directly to DNA/RNA samples without prestaining the gel.
• Integrated 10X loading dye: No separate loading dye is required.
• Low background fluorescence: Avoids staining the entire gel matrix, helping improve contrast between nucleic acid bands and the surrounding background.
• Supports downstream applications: According to the manufacturer's application data, the stain can support improved DNA recovery and downstream applications such as cloning, sequencing, and PCR.
• Alternative to ethidium bromide: The manufacturer describes the product as non-carcinogenic and less mutagenic than EtBr.
• Optimized for LED illumination: Provides strong fluorescence signals with blue LED and blue/green LED systems.

Key Differentiator

With MIDORI Green Direct, simply mix the stain with the DNA/RNA sample or DNA marker at the recommended ratio, load the sample, and perform electrophoresis. There is no need to add the stain to the agarose gel or running buffer.

2. Technical Specifications

3. Applications and Protocols

3.1. Applications

MIDORI Green Direct is suitable for molecular biology laboratories using agarose gel electrophoresis for:

• Analysis of PCR products and amplified DNA.
• Analysis of DNA following restriction digestion.
• DNA marker visualization and DNA fragment size assessment.
• RNA detection in agarose gels.
• DNA recovery from gels for downstream applications such as cloning, sequencing, and PCR.
• Simplifying nucleic acid staining workflows and reducing sample preparation steps before electrophoresis.

3.2. Recommended Procedure

According to NIPPON Genetics EUROPE's instructions:

1. Prepare the agarose solution and dissolve the agarose completely.
2. Mix the agarose solution gently, avoiding the formation of air bubbles.
3. Allow the solution to cool to approximately 60–70°C before casting the gel.
4. Mix DNA/RNA samples or DNA markers with MIDORI Green Direct at a 1:10 ratio (stain).
5. Load the samples into the gel wells and perform electrophoresis using an appropriate protocol.
6. Visualize and document the results using a gel documentation system. Blue LED or blue/green LED illumination is recommended for optimal signals; UV can also be used if necessary.

Important: Do not use more than 0.5 µL of MIDORI Green Direct per sample, even if this means the actual mixing ratio differs from 1:10. Consult the manual to confirm the appropriate volume for the sample amount and electrophoresis conditions.

3.3. Benefits for Downstream Applications

Some nucleic acid stains that intercalate into DNA can interfere with enzymes used in subsequent applications. According to the manufacturer, MIDORI Green Direct binds to the DNA backbone and was developed to support downstream applications such as cloning, sequencing, and PCR following DNA recovery from agarose gels.

NIPPON Genetics EUROPE provides comparative transformation-efficiency data from a specific application experiment. These results should be interpreted within the conditions of the manufacturer's experiment and should not be regarded as a performance guarantee for every sample or workflow.

4. Compatibility and Technical Considerations

4.1. Compatibility

MIDORI Green Direct is optimized for direct sample staining. It should not be used for in-gel staining or poststaining in the same way as MIDORI Green Advance, as the two products are designed for different staining methods.

4.2. Safety and Storage

According to the manufacturer, MIDORI Green Direct was developed as a safer alternative to ethidium bromide, with safety assessments addressing cytotoxicity, mutagenicity, and permeation through cell membranes or gloves.

These claims do not replace the need for appropriate laboratory safety practices.

• Read the SDS before use.
• Wear appropriate gloves and personal protective equipment.
• Avoid direct contact with the stain and used staining solutions.
• Store at 4°C and do not freeze, following the manufacturer's instructions.
• Dispose of waste according to the SDS and laboratory procedures.

4.3. Practical Recommendations

• MIDORI Green Direct already contains loading dye; a separate loading dye is unnecessary when the product is used as directed.
• Do not add MIDORI Green Direct to the agarose gel or running buffer.
• Blue LED and blue/green LED illumination are preferred for optimal fluorescence signals.
• When switching from ethidium bromide or another nucleic acid stain, reassess the illumination system and imaging settings.
• Do not directly substitute the MIDORI Green Advance protocol for MIDORI Green Direct, as the products differ in concentration and intended use.

Contact Us for Technical Support and Quotations

PACIFICLAB – Laboratory reagents and molecular biology solutions.

Contact PacificLab for technical consultation and assistance in selecting MIDORI Green Direct for your electrophoresis system, staining workflow, and analytical requirements.

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