Individual Transposition – Nuclei Input Calculator
Targeted Nuclei Recovery from Individual Transposition followed by GEM Generation: 500-20,000
Determine Nuclei Stock and Diluted Nuclei Buffer* volumes (total 15 μl) for the Transposition Mix using this guidance:
-
Based on the starting Nuclei Stock Concentration, locate the corresponding row in the table. Identify the cell with pre-calculated volumes that align with the Targeted Nuclei Recovery. These values already account for a Recovery Efficiency Factor of 1.41 from GEM Generation.
-
For concentrations not listed in the table, use the formula provided after the tables 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.
Individual Transposition — Formula for Nuclei Stock & Diluted Nuclei Buffer volumes
|
Nuclei Stock Volume (μl) |
= Targeted Nuclei Recov. (GEM Gen.) x 1.41 (Recovery Efficiency Factor-GEM Gen.) | ||
| Nuclei Stock Concentration (nuclei/μl) | |||
| Diluted Nuclei Buffer Vol. (μl) | = 15 μl - Nuclei Stock Volume μl |
Example Calculation
|
Nuclei Stock Volume (μl) |
= 20000 X 1.41 | = 3.5 μl | |
| 8000 | |||
| Diluted Nuclei Buffer Vol. (μl) | = 15 μl - 3.5 μl | = 11.5 μl |