The following is an example of data generated from a CRISPRtest experiment in MDA-MB-231 cells.

Flow Cytometry Scatter

Example of Flow Cytometry Scatter generated from a CRISPRtest experiment in MDA-MB-231 cells

Figure. CRISPRtest experimental data, showing flow cytometry scatter plot analysis of GFP and RFP expressing cells at Day 10 in (A) Parental and (B) Cas9 expressing MDA-MB-231 cells. The percentages of RFP-positive cells calculated in the GFP- RFP+ quadrant Q1 and GFP-positive cells in the GFP+ RFP- quadrant Q3 are used to calculate the percent knockout in the table below.

Calculations

Calculations Parental Cells Day10 Cas9 Cells Days 10
%RFP 8.29% 14.3%
%GFP 11.9% 0.42%
GFP:RFP = 11.9 / 8.29 = 1.44 = 0.42 / 14.3 = 0.03

Calculate %KO using [(GFP:RFP)Cas9 / (GFP:RFP)Parental] as follows:

1 – 1 – (0.03 / 1.44) = 98%

Last modified: Jun 17, 2020

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