The following is an example of data generated from a CRISPRtest_Blue experiment in MDA-MB-231 cells.
Flow Cytometry Scatter
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 Day 10 | Cas9 Cells Day 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 – (0.05 / 1.42) = 96%
Last modified:
15 March 2024
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