Figure 3.
Transformation efficiency and fidelity. (A) Transformation efficiencies for the in vivo assembly of 100 and 50 kb synthetic chromosomes. Both neochromosomes have been assembled with 2.5 kb (dark blue), 5 kb (lighter blue) and 10 kb (lightest blue) fragments. The number above each bar indicates the number of assembled fragments. Data were normalized to 108 cells using controls on YPD plates, performed in biological duplicates (replicate 1 and 2) in technical triplicates for replicate 1 and technical duplicates for replicate 2. The asterisks indicates that transformation with 5 kb fragments resulted in a significantly different transformation efficiency compared to both neighbouring efficiencies (two-tailed paired homoscedastic t-test P< 0.05). (B) Assembly fidelity. In vivo assembly of the 100 kb NeoChr12 from 43 fragments. Eleven colonies that grew on selective medium were tested for mRuby2 and mTurquoise2 fluorescence. Neochromosomes were subsequently screened based on their size on CHEF gel and lastly by Illumina sequencing. The percentage of correct colonies for each screening round is indicated.