Microbiology

21 days ago

This is a quick run-through of the protocol to make dilution spot plates for S. cerevisiae. This assay is great for identifying broad fitness defects but isn't quantitative. Note that temperatures for temperature sensitive strains will need adjustment from the standard 30°C. Even if you aren't aware of any temperature-rela...

about 2 months ago

Influenza A viruses, the culprits of respiratory illness in humans, are a complex and multifaceted group of viruses. Among the numerous subtypes of Influenza A, H1N1 and H3N2 with a notorious history of causing seasonal epidemics and pandemics, demand special attention. Influenza A H1N1, also known as the swine flu, is a f...

2 months ago

For those of y'all who couldn't attend the AMA... here's the recording of our salon session with Sebastian Cocioba!! Listen to hear the origin story and other tidbits from our favorite independent researcher and amateur biologist :) https://the-fancy-labcoat-guild.simplecast.com/episodes/small-thoughtful-science-a-live-ama-...

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...

4 months ago

Hi folks, I found a nice overview over the strength of various BioBricks promoters https://jbioleng.biomedcentral.com/articles/10.1186/1754-1611-3-4 I'm especially interested in finding a constitutive promoter (family) so I can test a range of promoters with different known strengths, for example the BioBricks J23100 prom...

7 months ago

Hello filamentous friends! Over the past few months, I've been troubleshooting growing Trichophyton rubrum, and attempting to get various strains to sporulate on solid media. In our lab, we have some experience with Aspergillus fumigatus but had never really struggled to get a strain to undergo sporulation. So, when I star...

8 months ago

Hi everyone, I've received a custom oligo pool from Twist Bioscience and they provided 70ng of it. We talked to one of the reps and they said not to resuspend it to any less than 10ng/μL, which means I'll be adding less than 7μL of buffer, which is obviously not a lot. I was wondering if anyone had any practical tips for h...

8 months ago

First of all, @Michael Paulsmeyer had a great post a while back on how to get as much DNA from a standard miniprep as possible (https://www.scifind.net/methods/efficient-miniprep-spin-column-k). The variance in my personal results from minipreps is large, though. To make sure I always get a high concentration of DNA, I use ...

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 ...

Dilution spot assay for yeast

Dilution spot assay for yeast

Influenza A: Unraveling the Mystery of H1N1 and H3N2

Influenza A: Unraveling the Mystery of H1N1 and H3N2

Sebastian Cocioba AMA recording on March 3rd 2023 for the Scifind Salon

Sebastian Cocioba AMA recording on March 3rd 2023 for the Scifind Salon

"DIY" Electroporation of NEB Turbo Using "Electropen"

"DIY" Electroporation of NEB Turbo Using "Electropen"

Any broad comparisons of promoter strengths?

Any broad comparisons of promoter strengths?

Troubleshooting Sporulation for Filamentous Fungi

Troubleshooting Sporulation for Filamentous Fungi

Resuspending Twist Oligo Pools?

Resuspending Twist Oligo Pools?

Miraprep: >30μg of Plasmid DNA Using a Miniprep Kit Every Time

Miraprep: >30μg of Plasmid DNA Using a Miniprep Kit Every Time

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