Simple Electroporation of Chlamydomonas reinhardtii Strains with an Intact Cell Wall
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Abstract
Chlamydomonas reinhardtii is a model green alga extensively used to study photosynthesis and cilia using molecular biology and genetics. Electroporation is a very common technique to transform DNA into the nuclear genome, which is essential to generate mutant collections and express transgenes. Here, we describe a simple, fast, and efficient protocol to transform strains with an intact cell wall. It achieves a good transformation efficiency without cell wall digestion or use of commercial kits and is compatible with the widely available Gene Pulser electroporation system.
Key features
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High transformation efficiency of Chlamydomonas reinhardtii strains with an intact cell wall.
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Faster than currently available electroporation protocols.
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Faster than currently available electroporation protocols.
This is one of the most compelling aspects of the manuscript to me. Gaining broad adoption really depends on lowering costs, reducing failure points, reducing the need for specialized equipment, and, importantly, speed.
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he transformation efficiency of our protocol was estimated at 13.3 x 103 transformants/μg of DNA or 134 transformant/106 cells, and the one of the ‘Crozet’ protocol was of 7.0 x 103 transformants/μg of DNA or 70 transformants/106 cells.
Are these efficiencies the results of a single experiment or averages of multiple experiments? How much experiment to experiment variation would you expect to see?
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Does not work with true cell-wall-less strains like UVM4
I think it is interesting that this method is incompatible with cell-wall-less strains. Do you know why that is?
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tested strains: CC-4051 (4A-), CC-5101 (T222+), CC-4425 (D66), CC-124, CC-4533, CC-5325
You have listed 6 strains as being tested, but only have data for 4A-. Is data for the remaining strains going to be included as supplemental data in the final pub? Are there any differences in transformation efficiency between strains?
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