We recommend starting with total RNA (DNA-free). For detailed sample submission requirements please visit our Sample Submission Guidelines.
To successfully extract RNA for RNA sequencing please follow these guidelines:
- Use Quality Reagents: Choose high-quality reagents and kits specifically designed for RNA extraction to ensure maximum yield and purity.
- Tissue or Cell Handling:
- If using tissue, immediately snap-freeze or store it in RNA preservation solutions (like RNAlater) to prevent RNA degradation.
- For cell cultures, process samples promptly and keep them on ice until extraction.
- Homogenization:
- Homogenize the samples thoroughly to ensure complete lysis of cells and release of RNA.
- Follow Protocols Carefully: Adhere strictly to the manufacturer's protocol of the RNA extraction kit. Pay close attention to:
- Sample input amount
- Lysis buffer volume
- Centrifugation times and speeds
- Avoid RNases:
- Use RNase-free tubes, pipette tips, and reagents. Wear gloves and use sterile equipment to minimize contamination.
- Check RNA Quality and Quantity:
- Use a spectrophotometer (e.g., NanoDrop) and a bioanalyzer / Fragment Analyzer to assess RNA purity (A260/A280 ratio) and integrity (RNA Integrity Number, RIN).
- Storage:
- Store extracted RNA at -80°C for long-term preservation. For short-term storage, RNA can be kept at -20°C.
- Prepare for Library Construction:
- Ensure that the RNA is free of contaminants (like DNA and proteins) and within the required concentration range for downstream applications.