Transformation Products for Algal Expression
MAX Efficiency® Transformation Reagent for Algae is a buffer that enhances transformation efficiency for multiple strains of Chlamydomonas species. One of the biggest hurdles in research and development with Chlamydomonas has been the introduction of exogenous DNA into Chlamydomonas strains due to rigid cell walls. Methods such as glass bead agitation, electroporation, microparticle bombardment are available but often result in low transformation efficiency. MAX Efficiency® Transformation Reagent increases permeability of the Chlamydomonas cell wall and facilitates increased delivery of DNA into the cell nucleus by electroporation. Simply grow the Chlamydomonas cells to early log phase, harvest by centrifugation, and wash twice with MAX Efficiency® Transformation Reagent prior to electroporation. To date, we have seen a >200-fold increase in transformation efficiency, over previously recommended conditions in our GeneArt® Chlamydomonas Engineering Kit and GeneArt® Chlamydomonas TOPO® Engineering Kit, in 10 different Chlamydomonas strains, including wild type and mutants, using circular or linear DNA as well as PCR fragments.
- Obtain >1,000 transformants/μg DNA for cell wall(+) strains
- Obtain >100 transformants/μg DNA for cell wall(–) strains
- Transform circular or linear DNA or PCR fragments
Eight different strains of Chlamydomonas reinhardtii, CC1690, CC2935, CC1009, CC4414, CC118, CC536, WT137c, CC3395 wall(–), were transformed using MAX Efficiency® Transformation Reagent for Algae and with TAP/sucrose reagent by electroporation. These data show MAX Efficiency® Transformation Reagent for Algae produced a >1000-fold increase in the number of colonies compared to that obtained using TAP/sucrose media for the culture of C. reinhardtii WT137c.