* Valid when your positive control sgRNA targeting RELA gives an editing efficiency of 50% or greater.
|4 x 600 pmol (20 µg)||Target-specific sgRNA, modified|
|1 x 600 pmol (20 µg)||Human RELA sgRNA, modified (positive control)|
|1 x 20 µL||Human RELA Primer Mix (10 µM)|
|1 x 20 µL||Human RELA Sequencing Primer (10 µM)|
|1.5 mL||Nuclease-free Tris-EDTA Buffer|
|1.5 mL||Nuclease-free Water|
|1 x 300 pmol||Cas9 2NLS Nuclease|
Four guide RNAs were designed per gene to knockout 9 kinases across the human genome. These were constructed as chemically modified synthetic sgRNA and electroporated into either HEK293 cells or Jurkat cells. The DNA from the resulting pool of cells was analyzed by TIDE analysis, demonstrating a high editing efficiency across the board with at least one guide per gene for every gene tested, giving greater than 50% editing efficiency (red line denotes 50% editing efficiency). This demonstrates the high efficiency with which guides designed and constructed within the Gene Knockout Kit perform.