Bacteria

2 months ago

Actinomycetes are known to be difficult to grow in micro-scale fermentations due to the aggregation of their pellets into "clumps", a common issue that leads to a lack of oxygenation in such a small space as a 96 or 24-well plate (suffocation if you will). So scientists are often afraid to scale down their fermentations, in...

4 months ago

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...

5 months ago

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...

8 months ago

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

9 months ago

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...

9 months ago

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 ...

9 months ago

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 ...

Scaling down fermentations of Actinomycetes

Scaling down fermentations of Actinomycetes

"DIY" Electroporation of NEB Turbo Using "Electropen"

"DIY" Electroporation of NEB Turbo Using "Electropen"

Help! My Tissue Culture Keeps Getting Contaminated! 😱

Help! My Tissue Culture Keeps Getting Contaminated! 😱

Broth Dilution Method (How to determine the MIC)

Broth Dilution Method (How to determine the MIC)

Why my KB Agar went cloudy and why you should check your growth medium reagents.

Why my KB Agar went cloudy and why you should check your growth medium reagents.

'You vs. the Guy She Tells You Not to Worry About': Heat-Shock vs. Electroporation

'You vs. the Guy She Tells You Not to Worry About': Heat-Shock vs. Electroporation

Efficient miniprep spin-column kit workflow for high plasmid yields

Efficient miniprep spin-column kit workflow for high plasmid yields