Fig. 5.

Synthesis of green and blue variants of EGFP using the SpeedyGenes method. (A) Oligonucleotides 6 and 12, which encoded the variant codon sequences, were ‘spiked’ into the final OE-PCR at a concentration of 6 nM each to assemble the full-length sequence. (B) In situ expression of these amplicons in E. coli yielded colonies with green fluorescence under blue light (EGFP, upper image) and/or blue fluorescence under UV light (EBFP, lower image). (C) Selected colonies from this plate were induced in liquid culture and the expression of EGFP (left-most three tubes) and EBFP (right-hand three tubes) was observed under UV light.