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Table 4 Comparison of expression library complexities provided by different combinations of linear and non-linear LR substrates using wt NTAIL sequence.

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

Donor

Acceptor construct

Number of clones

wt NTAIL-pDONR1

Non-linearized pNGG

0.3 × 107

wt NTAIL-pDONR1

SalI-linearized pNGG

0.4 × 107

wt NTAIL-PCR2

Non-linearized pNGG

1.6 × 107

wt NTAIL-PCR2

SalI-linearized pNGG

0.25 × 107

  1. The number of E. coli colonies provided by various combinations of 100 fmoles of donor and of 25 fmoles of acceptor construct in a 5 μl LR reaction is reported. T7pRos E. coli cells were electroporated and spread on ACplates
  2. 1wt NTAIL coding sequence borne by pDONR. 2wt NTAIL coding sequence amplified using att L1 and att L2 primers
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Microbial Cell Factories

ISSN: 1475-2859

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