Molecular Virology Support Core: qPCR Sample Submission Guidelines
The Molecular Virology Support Core provides specialized services for quantitative Polymerase Chain Reaction (qPCR) analysis to support research projects across the institution. To ensure the highest quality of data and experimental reproducibility, all researchers must adhere to the following sample submission guidelines. Please review these instructions carefully before preparing your samples.
1. General Preparation Requirements
Success in qPCR is highly dependent on the quality and integrity of the input nucleic acids. All samples must meet the following criteria:
- Purity: Ensure that your DNA or RNA samples are free from contaminants such as proteins, salts, ethanol, or phenol. 260/280 ratios should ideally be between 1.8 and 2.0 for DNA, and ~2.0 for RNA.
- Quantification: Samples should be quantified using a fluorometric method (e.g., Qubit) rather than spectrophotometric methods (e.g., NanoDrop) for greater accuracy.
- Integrity: RNA samples must be checked for degradation. Use an Agilent Bioanalyzer or similar capillary electrophoresis system to ensure the RNA Integrity Number (RIN) is acceptable for your experimental goals.
2. Sample Vessel Specifications
Proper containment prevents evaporation and cross-contamination. Please follow these storage standards:
- Use only certified DNA/RNA-free, thin-walled PCR tubes or 96-well plates.
- For plate submissions, ensure that a high-quality optical adhesive film is used to seal the wells securely.
- Clearly label the tube side or the plate skirt with a unique identifier. Do not label the caps alone, as they may be switched during processing.
3. Submission Documentation
Every submission must be accompanied by a completed qPCR Sample Submission Form. This form includes critical metadata required for downstream data analysis:
- Sample names and biological replicates.
- Target gene name and sequence information (if using custom primers).
- Expected concentration (ng/L) and total volume provided.
- Buffer composition (e.g., TE buffer, Nuclease-free water).
- Information regarding potential inhibitors present in the samples.
4. Shipping and Storage Standards
Maintaining the cold chain is essential for preventing nucleic acid degradation.
| Sample Type | Storage/Shipping Requirement |
|---|---|
| DNA | Shipped on dry ice or cold packs; stable at 4C for short term. |
| RNA | Must be shipped on dry ice and stored at -80C. |
| cDNA | Shipped on dry ice or cold packs. |
5. Quality Control Policy
The Core performs an initial Quality Control (QC) check upon receipt of samples. If a sample fails to meet the minimum concentration or purity requirements, the core will notify the Principal Investigator immediately. If the user decides to proceed with sub-optimal samples, the Core is not responsible for suboptimal results, including high Cq values or flat amplification curves.
6. Data Return and Confidentiality
Results will be provided in a standard format, including raw amplification plots, Cq values, and melt curve data. Data is held in a secure database and is accessible only to the specific research team associated with the project. After the completion of the project, samples are kept for 30 days before being discarded unless other arrangements have been made in writing.
If you have any questions regarding your specific experimental design or require troubleshooting assistance, please contact the Molecular Virology Support Core staff at least one week prior to your planned submission date.
