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. 2021 Nov 9;16(11):e0259812. doi: 10.1371/journal.pone.0259812

Fig 2. Example of transfection plate layout.

Fig 2

Experiments where (A) optimal ratio of gRNA to CRISPR/Cas protein required for precomplexing will be determined, (B) the optimal amount of recombinant CRISPR/Cas will be determined (if we assume the optimal ratio will be 1:1) and (C) optimization of transfection reagent amount (1×, 2× and 3×) used for complex formation (if we assume the optimal CRISPR/Cas amount is 0.6 pmol per well). Wells named “T.R.” will contain transfection reagent only. White wells will hold non-transfected cells used as non-treated controls for the cell viability assay. The crossed-out wells represent empty wells used for holding the PrestoBlue™ Cell Viability Reagent staining blank. Grey wells will be filled with PBS to avoid edge effect. All experiments will be performed in triplicate, 4 gRNAs will be used for RNP formation (AAVS1, CDK4 and HPRT1 and non-targeting control gRNA).