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Table 5 Comparison of expression library complexities provided by different combinations of epPCR products and of non-linearized acceptor constructs

From: One-step generation of error-prone PCR libraries using Gateway® technology

epPCR product Non-linearized acceptor construct Number of clones
NTAIL1 10 fmoles pNGG4 2.8 × 107
NTAIL1 100 fmoles pNGG4 3.25 × 107
LadS2 10 fmoles pTH315 6.65 × 107
LadS2 100 fmoles pTH315 3.85 × 107
PA30593 42 fmoles pTH315 1 × 107
  1. The number of E. coli colonies provided by various combinations of epPCR product and of 25 fmoles of acceptor constructs in 5 μl of LR reaction is reported. T7pRos E. coli cells were electroporated and spread on ACplates
  2. 1NTAIL PCR product is 661 bp. 2LadS PCR product is 696 bp. 3PA3059 PCR product is 1,780 bp. These three PCR product lengths include the att L1 and att L2 recombination sites. 4pNGG is 8,012 bp before LR recombination. 5pTH31 is 8,306 bp before LR recombination.