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. 2020 Jul 2;10:10894. doi: 10.1038/s41598-020-67791-z

Figure 1.

Figure 1

Graphical representation of the plasmid assembly method using an amplicon containing 96 unique barcodes amplified from a library of pJazz Knockout vectors2. (A) Amplicon assembly of all 96 barcodes into the pCC1-hDHFR transfection vector was performed in a single Gibson reaction, and the bacterial transformation was used to directly seed a large overnight culture for midiprep plasmid preparation. This starting pool was examined by Bar-Seq to confirm representation of barcodes (see Fig. 2). (B) Pools of 94 constructs were used as DNA input for transfection. (C) Parasite genomic DNA was extracted from both bulk cultures and individual clones for barcode amplification from episomes for Next Generation Sequencing and barcode quantification.