Plan transfection master mix volumes for DNA or RNA experiments. Estimate total nucleic acid and transfection reagent volumes per well and for all wells, including overage for pipetting loss.
Enter your wells, DNA amount, and reagent ratio to plan your transfection master mix.
A transfection master mix is a pre-mixed solution containing nucleic acids (DNA plasmids, siRNA, mRNA, etc.) and transfection reagent that can be distributed across multiple wells or vessels. Preparing a master mix ensures consistent reagent ratios across all samples and reduces pipetting errors compared to preparing each well individually.
Effective Wells = Wells × (1 + Overage% / 100)Total Mass (µg) = Mass per Well × Effective WellsStock Volume (µL) = Total Mass / Stock ConcentrationReagent Volume (µL) = Total Mass × Reagent RatioThe amount of nucleic acid per well and the reagent-to-DNA ratio are critical parameters that affect transfection efficiency and cell viability:
| Plate Format | Typical DNA (µg/well) | Cell Count (approx.) |
|---|---|---|
| 6-well | 2.0 – 4.0 | 3–5 × 10⁵ |
| 12-well | 0.8 – 1.5 | 1–2 × 10⁵ |
| 24-well | 0.4 – 0.8 | 5 × 10⁴ – 1 × 10⁵ |
| 96-well | 0.05 – 0.2 | 1–3 × 10⁴ |
Note: These are general guidelines. Actual amounts depend on cell type, reagent, and experimental goals.
When preparing a master mix for multiple wells, some volume is inevitably lost due to:
A 10% overage is commonly used, meaning you prepare enough mix for 10% more wells than you actually need. For example, for 10 wells with 10% overage, you would prepare enough for 11 effective wells.
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Plan master mixes efficiently and reduce pipetting errors in your molecular biology workflows
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