This is a method for low-cost electrotransformation of electrocompetent NEB turbo E coli cells using a homemade electroporation apparatus. The original paper upon which this method is based can be found here: https://doi.org/10.1371/journal.pbio.3000589 My method for preparing electrocompetent E coli can be found here: h...
I am working with carrots, which are, in general, very easy to culture. Just pop them on MS media with 2,4-D and they will produce callus within a month. My only problem is that my carrots seem to have a LOT of endophytic bacteria that take over very quickly. What can I do to suppress or kill these bacteria? I plan on using...
Hey guys, I have visually described the Broth Microdilution assay, a method that is commonly used in antibiotic research to determine the minimum inhibitory concentration (MIC) of a compound in bacteria: https://www.youtube.com/watch?v=nxcU9_XyXsE Best Henrik
I want to talk about preparing basic microbiological growth media and things that can go wrong when swapping reagents. Weird cloudy KB Agar A while ago, I was preparing KB Agar: Recipe for 1L KB Agar 10g Glycerol 20g Peptone 1.5g K2HPO4 trihydrate 12g Agar powder 5 mL sterile 30% MgSO4 solution (to be added after autocla...
I compared the results from identical cloning protocols preceding either a heat-shock or electroporation bacterial transformation into NEB competent E. coli cells (chemically competent or electrocompetent) and found that the electroporation transformation consistently resulted in thousands more colonies than the heat-shock ...
Here is my workflow for high plasmid yields from cultures of DH5α E. coli. I use the GeneJet Plasmid Miniprep kit from Thermo (#K0502), but most spin column kits work the same. Generally I have to extract plasmids from numerous cultures simultaneously, so this protocol also gives guidance on the timing of each step. Prick ...
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