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

Flow Cytometry Scatter

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

Figure. CRISPRtest_Blue experimental data, showing flow cytometry scatter plot analysis of BFP and RFP expressing cells at Day 10 in (A) Parental and (B) Cas9-
expressing 293T cells. The percentages of RFP-positive cells calculated in the BFP- RFP+ quadrant Q3 and BFP-positive cells in the BFP+ RFP- quadrant Q1 are used to calculate the percent knockout in the table below.

Calculations

Calculations Parental Cells Day10 Cas9 Cells Days 10
%RFP 12.9% 19.5%
%BFP 18.3% 1.15%
BFP:RFP = 18.3 / 12.9 = 1.44 = 1.15 / 19.5 = 0.06

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

1 – 1 – (0.05 / 1.42) = 96%

Last modified: Jun 17, 2020

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