Two-Stage Pooling Protocol
- Stage 1: For each group, pipette the suggested volumes from each library into a separate tube to create group subpools.
- Stage 2: From each group subpool, pipette the 'Take Volume (Stage 2)' into the final pool tube.
- Final step: Add the required water volume to reach the target final volume.
Input File Format
Upload a CSV file with the following columns:
- Sample: Sample identifier (string)
- Ct: Cycle threshold at 2/3 max fluorescence (numeric)
- nM: Library concentration in nM (numeric)
Ct-to-UMI Model
The default model is:
N_i = 8.44 × 10^7 × e^(-0.555 × Ct_i)
You can customize this expression. Requirements:
- Use 'Ct' as the variable name
- Expression must be vectorized (work on vectors)
- Must return positive numeric values
Final pool volume cap (local modification)
If Group 3 exists, the app will cap your requested final pool volume so that:
Final_Pool_Volume_Used <= Subpool_Volume_Stage1_uL (Group 3)
This treats Group 3 Stage 1 subpool volume as a limiting factor.