GenBuilder™ DNA Assembly kit- a new look at synthetic biology
GenScript’s GenBuilder™ DNA Assembly technology helps you easily perform modifications to your DNA. The applications range from expressing synthetic pathways in living cells to analyzing protein function using saturation mutagenesis.
Case study #1 GenBuilder™ DNA Assembly to assemble the violacein biosynthetic pathway (8kb) into pUC57
Figure 1. Pathway assembly using GenBuilder™. The violacein biosynthetic pathway (8kb) was assembled using the GenBuilder™ DNA Assembly. A. The pathway consists of five genes, which were assembled as six separate fragments into the pUC57 vector. B. The pathway was correctly assembled, and was functional in producing the purple violacein pigment in DH10B cells. PCR and sequencing analysis further confirmed the seamless DNA assembly.
Case study #2 GenBuilder™ DNA Assembly for using ssDNA as a bridge to introduce codon saturation mutagenesis for directed evolution
Figure 2. Using ssDNA as a bridge to create saturation mutagenesis for directed evolution. The RFP gene consisting of two fragments was mutated with ssDNA fragments. A. Q66 was targeted for NKK saturation mutagenesis in between 426 bp and 661 bp RFP sub-fragments. B. Codons 194, 195, and 196 were targeted for NKK saturation mutagenesis in between 810 bp and 271 bp RFP sub-fragments. The DNA fragments were assembled into pUC57 vector. C. Both reactions produced hundreds of colonies in different colors, indicating the successful introduction of various mutations in RFP. 24 clones were randomly picked for sequencing of the 194-196 saturation mutagenesis sample and found >92% of codon diversity