gBlocks Gene Fragments were assembled using the Gibson Assembly™ Method to generate some of the plasmid constructs involved in refactoring a N-demethylation operon that provides a caffeine degradation pathway from Pseudomonas putida CBB5 to E. Coli, an organism more amenable to genetic engineering and industrial applications. The resulting cell lines required caffeine to survive and their growth yield could serve as a biosensor to precisely measure caffeine concentrations, for example, in sodas and energy drinks. This work was part of a 2012 undergraduate iGEM project sponsored by IDT.
gBlocks Gene Fragments were used to generate expression constructs for optimized sgRNAs, that were then used to target a nuclease-deficient, Cas9-GFP chimeric protein in chromatin imaging experiments.
A codon-optimized Cas9 was created by ordering a series of overlapping gBlocks Gene Fragments, and assembled using the Gibson Assembly™ Method, and inserted into the vector pCFJ601. In addition, a single gBlocks Gene Fragment was inserted downstream of the Cas9 sequence, containing a U6 promoter, and the necessary elements for sgRNA function.
gBlocks Gene Fragments are used to generate constructs for yeast two-hybrid assays.
The authors have developed a multiplex PCR method that allows for quantitative analysis of T- and B- cell receptor diversity using next generation sequencing. gBlocks Gene Fragments are used as templates for optimization of the multiplex primer mix, a critical step for maintaining quantifiable differences and detecting low level transcripts.
Sets of hybridized, high-fidelity Ultramer Oligonucleotides were assembled into large constructs in a single step. Such constructs are available from IDT as gBlocks® Gene Fragments.