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 and the calculator 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  

Individual Transposition — Calculator for Nuclei Stock & Diluted Nuclei Buffer volumes

fixed value

minimum: 500
maximum: 20,000

minimum: 1,000
maximum: 12,000

μl

μl

Low transfer volume that may result in higher nuclei load variability
Volume would exceed the allowable buffer volume in each reaction

Numbers are rounded to the nearest decimal.