Scalable Transposition – Nuclei Input Calculator
Targeted Nuclei Recovery from Transposition: 20,000-75,000
Determine Nuclei Stock and Diluted Nuclei Buffer* volumes (total 15 μl) for the Transposition Mix using this guidance:
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Based on the starting Nuclei Stock Concentration, locate the corresponding row in the table. Identify the exact cell with pre-calculated volumes based on the Targeted Nuclei Recovery. These values already account for a Recovery Efficiency Factor of 1.0 from Transposition.
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For concentrations not listed in the table, use the formula provided after the table to calculate custom volumes.
Higher nuclei concentrations result in smaller pipetting volumes, which may increase variability.
*Use ONLY Diluted Nuclei Buffer (Dilute 20X Nuclei Buffer (PN-2000207) 1:20 in nuclease-free water) as specified in the previous page.
Scalable Transposition — Formula for Nuclei Stock & Diluted Nuclei Buffer volumes
|
Nuclei Stock Volume (μl) |
= Nuclei Input for Transp. x 1.0 (Recovery Efficiency Factor-Scal. Transp.) | ||
| Nuclei Stock Concentration (nuclei/μl) | |||
| Diluted Nuclei Buffer Vol. (μl) | = 15 μl - Nuclei Stock Volume μl |
After Scalable Transposition, nuclei concentration is determined and the transposed nuclei input volume for GEM Generation is adjusted based on Targeted Nuclei Recovery from GEMs, factoring in Recovery Efficiency Factor for GEM Generation (1.41), as specified in Transposed Nuclei Suspension Volume Calculator Table for GEM Generation
Example Calculation
|
Nuclei Stock Volume (μl) |
= 75000 X 1.0 | = 9.4 μl | |
| 8000 | |||
| Diluted Nuclei Buffer Vol. (μl) | = 15 μl - 9.4 μl | = 5.6 μl |