Standards plate planning for assay preparation

Worked example: four standards in duplicate

Start with a 2,000 µM stock. Set the highest standard to 1,000 µM and a two-fold series to obtain 1,000, 500, 250 and 125 µM. Allocate two 100 µL wells per standard and two blank wells.

For this example, use 200 µL minimum usable volume per standard, 20% overage and 0 µL additional dead volume. The required retained volume is 240 µL per standard. Preparation volumes also cover the transfer needed to make the next standard.

1. Review the preparation volumes

For S1, 225 µL of 2,000 µM stock plus 225 µL diluent gives 450 µL at 1,000 µM. Transfer 210 µL to S2, leaving 240 µL. The same balance applies down the series. Prepare 240 µL of the appropriate blank diluent separately.

2. Check the plate allocation

Standards occupy A1–A8; blanks occupy A9–A10. This illustrative map uses 10 of 96 wells. It does not allocate unknown samples or perform curve fitting.

3. Save the setup and keep the working

In BenchLine, save a valid setup for repeat work. Export the preparation table or plate map as CSV, or a supported calculation report with inputs, assumptions and warnings.

These downloads describe this website example. They are calculation-support illustrations, not a validated assay protocol or a substitute for checking your method.