Processing
IDT gBlocks® Gene Fragments provide a tool for easy gene construction or modification. Single fragments can be used to introduce multiple modifications into a sequence, or as genes substitutes (e.g., for short genes such as an immunoglobulin (Ig) domain) [1–2]. Several of these double-stranded DNA fragments of up to 3000 bp each can be quickly assembled using the Gibson Assembly™ method to generate genes or other constructs [3]. Thus you can re-create the elusive or damaged gene from your clone library, re-constitute a gene from an organism that is not readily accessible, or design and build a new function or chimera. Additional applications of gBlocks Gene Fragments include using them as template or substrates for enzymatic reactions (polymerases, methylases, etc.) or as standards for qPCR [4].
Gene construction for protein engineering
In one example, the Kirill Alexandrov laboratory at the Institute of Molecular Biology (IMB), University of Queensland (Brisbane, Australia) uses gBlocks Gene Fragments in their studies of intracellular vesicular transport and the role played by Rab GTPases in docking and fusion of intracellular membranes. The lab performs a lot of protein engineering and gBlocks Gene Fragments have provided them with rapid and cheap access to new types of functional and structural elements that they do not possess in their plasmid database. Thus, the researchers can construct novel biological modules and cascades to better understand the tethering complexes and transcription-regulating complexes critical to their system.
Easy assembly by molecular biology novices
The Alexandrov lab predominantly employs the Gibson Assembly method. They find it a very fast and robust technique that requires a minimal amount of skill and expertise. Dr Viktor Stein, a postdoctoral fellow in the lab (Figure 1), notes, “Students who are only in their third week in the lab are routinely assembling 3–4 DNA fragments right after they have mastered plasmid preps and PCRs.”
Fidelity of constructs
“The fidelity of gBlocks fragments is generally very good, especially for assembling synthetic DNA fragments less than 1 kb,” says Dr Stein. The group verifies cloned DNA assemblies by sequencing. Note that when assembling gBlocks Gene Fragments, IDT scientists recommend sequencing twice as many clones as the number of gBlocks fragments assembled (e.g., when assembling 3 fragments, you would sequence 6 clones).
