Transformation of Chlamydomonas
One of the biggest hurdles in research and development with Chlamydomonas has been the introduction of exogenous DNA into Chlamydomonas strains, the difficulty being due to their rigid cell walls. Methods such as glass bead agitation, electroporation, and microparticle bombardment are available but often result in low transformation eﬃciency. Invitrogen™ MAX Eﬃciency™ Transformation Reagent for Algae is a buffer that enhances transformation efficiency for multiple strains of Chlamydomonas.
MAX Eﬃciency Transformation Reagent increases the permeability of the Chlamydomonas cell wall and facilitates increased delivery of DNA into the cell’s nucleus by electroporation. Simply grow the Chlamydomonas cells to early log phase, harvest by centrifugation, and wash twice with the MAX Eﬃciency Transformation Reagent prior to electroporation. We have seen substantial transformation efficiencies in ten 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 with circular or linear DNA or PCR fragments
Eight diﬀerent strains of Chlamydomonas reinhardtii (CC1690, CC2935, CC1009, CC4414, CC118, CC536, WT137c, and CC3395 wall(–)) were transformed by electroporation using MAX Eﬃciency Transformation Reagent for Algae for delivery followed by resuspension in TAP/sucrose reagent prior to plating for growth. These data show that the MAX Eﬃciency™ Transformation Reagent for Algae produced a >1,000-fold increase in the number of colonies compared to the numbers obtained using TAP/sucrose medium alone for the culture of C. reinhardtii WT137c.
For Research Use Only. Not for use in diagnostic procedures.