This workflow connects copy number conversion, dilution planning, and final qPCR setup into one sequence so the standard curve can be defended and repeated later.
Steps
- Confirm the identity and length of the standard molecule.
- Convert the measured concentration into copies per unit volume.
- Decide the target copy range and number of points for the curve.
- Plan the dilution series with enough material for replicates, repeats, and dead volume.
- Label the full series before starting any transfers.
- Prepare the qPCR master mix and reserve enough wells for controls and standards.
- Record the final copy range, transfer scheme, and any re-quantification notes.
Control points
- A wrong molecule length will poison the entire standard curve.
- A perfect dilution pattern is still weak if the starting concentration estimate is stale.
- Replicate volume must be protected before the first transfer is made.
Use these tools
- DNA/RNA Copy Number
- Serial Dilution Planner
- PCR/qPCR Master Mix
Tips
- Keep one line in the notebook with the molecule length, source batch, and target range.
- Prepare standards first so the reaction layout is known before master mix aliquoting.
- When possible, keep a frozen parent stock separate from the working dilution series.