PoolMate

Pool libraries into one tube. Enter your samples, then use the actions at the bottom: Calculate molarities (quick nM check) · Quick pool table (manual pooling, no robot files) · Generate worklists (full epMotion DNA + Buffer export).

Run metadata

Run name

Operator

Kit lot

Instrument / method

Notes

Create pool parameters

Platform

Loading preset

Pool calculation mode*

Target pool nM*Use when your SOP defines a final pool concentration.

Total final volume µL*

Layout*

Optional extra pooling parameters

Minimum transfer µL

Round transfer to µL

Dead volume / excess %

Source rack number

Destination rack number

Buffer rack numberUsually the same as the source rack. Only change it if your epMotion method has a dedicated buffer position on another rack (many fixed pooling scripts only use racks 1–2).

Buffer source position

Pool well

Low-volume handling

Pre-dilution final volume µL

Pre-dilution safety margin %

Tip policy

Max well capacity µL

Dilution rack number

Dilution starting well

These fields have safe defaults. Leaving this section closed does not block worklist generation.

Minimum for Generate Worklists: sample molarities plus pool mode, total final volume, and layout. Use nM mode for Illumina-style final concentration. Use fmol mode for ONT-style total load.

Quick pool method

What this does

Creates a simple manual pooling table from the Sample section without requiring platform presets, deck settings, destination racks, buffer positions, or epMotion worklist exports.

Minimum required: sample molarity inputs plus either fmol per sample or total fmol pool. This is for quick bench calculations, not full robot execution.

Quick pool target

Quick pool mode*

fmol per sample*

Round transfer to µL

Outputs transfer volumes only. Use Generate Worklists when you need epMotion files.

Samples

Shared fragment size (bp), optional

Default input mode

This form starts blank each time the file is opened. Click Save run to keep your work as a file you can reopen — unsaved entries are cleared on the next open.

#	Source well	Name / number	Input mode*	Conc reads ng/µL*	Size bp*	Direct nM*

Samples entered0

Mass-mode samples0

Overall avg concentration-

Concentration range-

* Minimum for Calculate Molarities: either concentration reads + size bp, or direct nM. Sample names and source wells help with tracking but are auto-filled if left blank. For replicate Qubit reads, enter values separated by commas, semicolons, or spaces. The overall average concentration below is calculated across mass-mode samples only.

Pool preview

Add samples or source wells to preview the layout.

Normalize — dilute each library to a target concentration (plate → plate)

Enter each library's well and concentration. PoolMate dilutes each one to the target concentration in a separate output plate and exports Normalization_DNA.csv + Normalization_Buffer.csv for your Eppendorf normalization script: source plate (Rack 1) → output plate (Rack 1), buffer from the reservoir at position "1". Output wells match the source wells.

Concentration unit*

Target concentration*

Final volume per well µL*

Min transfer µL

Round to µL

Well	Name	Concentration*

Target and each concentration must use the same unit. The target must be below every library's concentration (this dilutes, it can't concentrate).

Standalone dilution calculator

Dilute one sample or a whole list to a target concentration — completely independent of pooling. Enter each sample's stock concentration; every other parameter defaults to the shared values below but can be overridden per row. Serial steps appear automatically when one dilution can't reach the target within the well limit. C1V1 = C2V2; bench reference only, no robot file.

Unit

Default target conc*

Default final volume µL*

Default min transfer µL

Default max well/tube capacity µL

Default round to µL

#	Name	Stock conc*	Target	Final µL	Min µL	Max µL	Round µL

Independent of the pool above. Any blank cell falls back to the matching shared default above.

About the numbers

Molarity for dsDNA: nM = conc(ng/µL) × 10⁶ / (660 × size_bp). Direct nM can be entered when molarity has already been measured. Transfers are calculated as equal fmol per sample, then rounded to the selected increment and audited for post-rounding deviation. Low transfer volumes can be handled by warning only, increasing final pool volume, or producing a pre-dilution plan. In fmol mode, the tool derives target nM as total fmol divided by final volume.

PoolMate

What this does

Quick pool target

PoolMate - User Guide

1. Calculate Molarities

2. Quick Pool

3. Generate Worklists

Pooling vs Sample Sections

Plate Map

Output Files