Frequently Asked Questions

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Storage conditions vary by product. Specific storage requirements are listed on each product page.

Yes, COAs are available for download in the Knowledge Hub and with shipment.

A positive control helps validate your experimental system by confirming that your transfection, RNA extraction, and detection methods are working properly. Using a siRNA positive control with known and expected results ensures the reliability of your RNAi experiment.

Both controls serve distinct purposes:

• Positive controls (like validated siRNAs targeting GAPDH, ACTB, or GFP/EGFP) verify that your experimental system is working effectively
• Negative controls (non-specific siRNAs with universal or random sequences) demonstrate the specificity of your siRNA’s effect

While negative control groups should show similar results, some variation may occur. If you observe larger than expected deviations:

• Check the accuracy of your experimental procedures
• Consider that stress-response and immune-related genes may be particularly sensitive to external factors
• Note that these sensitive genes can show variable expression changes under experimental conditions

Common fluorescent dyes have the following specifications:

• FAM: absorption at 495nm, emission at 520nm
• Cy3: absorption at 550nm, emission at 565nm
• Cy5: absorption at 643nm, emission at 667nm

You can assess genOFF™ siRNA effects through three main methods:

• Measure mRNA levels using Realtime RT-PCR
• Detect protein levels using Western blot or ELISA
• Observe cell phenotype changes using high-resolution microscopy (based on your gene of interest’s function)

The effectiveness of genOFF™ siRNA can vary between different cell types due to several key factors:

1. Transfection Efficiency

• Different cell types have varying abilities to uptake genOFF™ siRNA
• Cell membrane composition and structure can affect siRNA entry
• Cell growth rates and density can influence transfection success

2. Target Gene Expression

• Baseline expression levels of the target gene may differ between cell types
• Regulatory mechanisms controlling gene expression vary across cell types
• Different splice variants may be present in different cell types

3. Additional Factors

• Metabolic state and cell cycle phase
• Presence of varying endogenous RNA interference machinery
• Different intracellular degradation rates of GenOFF™ siRNA

Optimization Tips

• Adjust transfection conditions for each cell type
• Validate knockdown efficiency using appropriate controls
• Consider using different delivery methods if necessary

Note: For optimal results with genOFF™ siRNA, please contact our technical support team for cell-specific transfection protocols.

RiboBio offers comprehensive siRNA design services for our genOFF™ products. Here’s how to get started:

1. What We Need From You:

• Gene name or Gene ID
• Species information
• Target sequence (optional)

2. Our Design Process:

• Advanced algorithm-based sequence selection
• Off-target analysis
• Secondary structure evaluation
• Multiple siRNA designs per target for optimal results

3. Design Service Features:

• Free design service
• Multiple targeting sequences provided
• Fast turnaround time

4. Ways to submit:

• Contact our customer service at support@ribobio.com
• Submit through our online portal

Generally, genOFF™ siRNA designed for one species may not work effectively in others, even for homologous genes.

Here’s why:

1. Sequence Differences

• While genes may be homologous across species, their exact sequences often differ
• Even small sequence variations can significantly impact siRNA effectiveness
• genOFF™ siRNA requires perfect sequence complementarity for optimal silencing

2. Key Limitations

• Most siRNAs cannot achieve cross-species gene silencing
• Even highly conserved gene regions may show species-specific variations
• Gene expression mechanisms can differ between species

3. Species-Specific Factors

• Different regulatory mechanisms
• Varying gene expression levels
• Species-specific splice variants
• Different cellular environments

Solutions We Offer:

1. Predesigned Validated genOFF™ siRNA
   • Ready-to-use validated siRNAs for common research targets
   • Available for multiple species
   • Experimentally validated for knockdown efficiency
   • Time-saving alternative to custom design
2. Custom Design Service
   • Specific design for your target species
   • Multiple sequence options
   • Optimized for your research needs

Recommendation: Check our catalog for predesigned validated genOFF™ siRNA first. If your target isn’t available, our design service can help you select the most effective sequences for your species of interest.

Contact our technical support team for species-specific genOFF™ siRNA designs to ensure optimal knockdown efficiency.

No, GenOFF™ siRNA is designed for room-temperature shipping while maintaining product stability:

1. Shipping Conditions

• Supplied as lyophilized powder
• Stable at room temperature during transport
• No cold chain shipping required
• Protected packaging ensures stability

2. Storage Requirements After Receipt

• Store at -20°C to -80°C
• Keep in a sealed container
• Protect from moisture
• Avoid repeated freeze-thaw cycles

3. Stability Features

• Lyophilization ensures product stability
• Maintains integrity during transportation
• Long shelf life when properly stored

Best Practices:

• Upon receipt, immediately transfer to -20°C or -80°C storage
• Follow resuspension protocol carefully
• Aliquot reconstituted siRNA to avoid repeated freeze-thaw

For detailed storage and handling instructions, please refer to the product manual or contact our technical support team.

No. genOFF™ siRNA lyophilized powder remains stable at room temperature for up to 2 weeks during shipping. For long-term storage after receipt, please store at -20°C to -80°C.

For optimal handling of genOFF™ siRNA:

1. Resuspension

• Centrifuge tube before opening
• Add RNase-free H2O or sterile ddH2O
• Prepare 20μM stock solution

2. Storage

• Store aliquots at -20°C to -80°C
• Avoid repeated freeze-thaw cycles

For detailed resuspension protocols, please refer to the product protocol or contact our technical support.

To maintain genOFF™ siRNA quality:

• Store in small aliquots after resuspension
• Limit freeze-thaw cycles to less than 5 times
• Avoid repeated thawing of the same tube

For optimal storage guidelines, refer to the product protocol or contact our technical support team.

No. GenOFF™ siRNA is pre-annealed during manufacturing and ready to use after resuspension.

There are several key factors that can affect the reproducibility of miRNA mimic experiments:

1. Spatial and temporal specificity: miRNA expression and function are highly dependent on cell type, developmental stage, and physiological conditions.

2. Technical considerations:

• Transfection efficiency variations between experiments
• Quality and concentration differences in miRNA mimics
• Cell culture conditions and cell passage number
• Timing of sample collection after transfection

Recommendations for better reproducibility

• Standardize transfection protocols and conditions
• Use multiple biological replicates
• Include appropriate controls
• Document detailed experimental procedures
• Validate results using alternative methods

While both micrOFF™ miRNA inhibitors and antagomirs are designed to suppress miRNA function, they have distinct characteristics that make them suitable for different applications:

micrOFF™ miRNA Inhibitors:

• Single-stranded RNA oligonucleotides with chemical modifications (2′-OMe)
• Shorter sequences (typically 21-23 nucleotides)
• Ideal for cell culture experiments due to:
  • Good cell membrane penetration
  • Cost-effective
  • High stability in cell culture conditions
• Limited in vivo stability

micrOFF™ miRNA Antagomirs:

• Cholesterol-conjugated single-stranded RNA analogs
• Additional chemical modifications (2′-OMe, phosphorothioate)
• Superior for in vivo applications due to:
  • Enhanced stability against degradation
  • Better tissue distribution
  • Longer lasting effects
  • Efficient cellular uptake without transfection reagents
• Higher cost compared to standard inhibitors

Selection Guidelines:

• For cell culture: Either option works well; inhibitors are more economical
• For animal studies: micrOFF™ Antagomirs are strongly recommended due to their enhanced in vivo stability and distribution
• For long-term studies: micrOFF™ Antagomirs provide more sustained effects

Ribobio also offers micrON™ miRNA agomir and micrON™ miRNA mimics as reliable tools for your miRNA research needs.