Are you trying to do transient or stable transformation? Nuclear or chloroplast integration? Is the genome of nucleus and chloroplast known for both species?
IIRC chlorella has a cell wall which prevents efficient ingress of just about anything. Protoplasting via cellulase may increase CaCl2 but I've read a few papers on chlorella a while back citing electroporation being the "bee's knees" but heck, electroporation can insert DNA into most living tissues so it should be a surefire way. If you do go the protoplast route, give polyethylene glycol (PEG) a chance. Its more gentle and less salty. Either way for transient expression you just need to get the plasmid in there, for actual stable integration...thats a whole different ball game.
test--On Wed, Dec 16, 2015 at 10:46 AM, Alex D <alexxxdean@gmail.com> wrote:Hello everyone,--I am currently working on a project which requires transfection of Chlorella Vulgaris and nannochloropsis. Plasmid size is approx, 6K bp, but I am more concerned about the most efficient and cost effective technique for transfection. Initial experiment will be carried with CaCl and later on by electroporation incase salt based method wont bring any results. But I wanted to try out sonification since it is in a way easier and cheaper to build/acquire and perhaps safer)).I am also working on developing more energy efficient containment for the culture to use minimal energy for shaking and molecule extraction methods, so anyways I dont have a PhD or anything I am a undergrad student but you can probably use some fancy words if you feel like. I am interested if anyone ever used multiple techniques and has any feedback on it which ever it is bad or not.Thanks in advance.
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