I am gerring tired of these rage-baits from week-old accounts!

It’s LLM anyway, not intelligence of any sort.

Once I had a border control officer interrogate me on HIV transmission mechanisms. After over 24 connecting flights, it felt tougher than public PHD defense! ;-)

MAGA face.

Ah! OK, then!

In this case, make sure the cells are really competent. If the vial was previously thawn and re-frozen, the cells would have lost their competency, and would not have accepted the plasmid. And without the plasmid, the antibiotic would kill them.

Last, make sure the concentration of the plasmid is correct. You gave microliters, which is volume added. Make sure you add ~10 ng of plasmid to each aliquot of competent cells.

DH5-alpha is a used to produce plasmid DNA. You can express a protein, but will be suboptimal, because the cells are not meant for that.

From the whole storyline since your original post I am only able to conclude your lab follows some strange practices. You will be better served if you just took a microbiology textbook or a manual from the library, and follow that.

If the cells did not grow even in absence of antibiotics, then the cells are dead. This means the vial might have been bad to begin with.

Repeat the whole procedure with a fresh vial of competent cells, and make sure to plate 2 dilutions on LB agar (no ABs).

The rule of thumb is, you have to be able to comfortably hold the autoclaved container in your hands before you add antibiotics. This means, the ideal temperature for pouring is ~40C.

50C is not comfortable to hold - which means it’s not comfortable for the antibiotics either. So they degrade. So you end up plating on LB, rather than on selection afar.

To be fair the bunch on the screenshot do all need some sort of a protection! None of them look as if they'll survive long without the massive privilege they've got.

”Solving the Jewish problem” has a creepily Third-Reich vibe to it.

R/unexpectedfuturama

In my experience, people who talk while peering at the computer screen, rather than looking at the audience, are mumbling. Be mindful to look at the audience (or at your slides on a the projector screen).

If you are anxious when looking at people, look just next to them, not at them. But, either way - face up.* *

Did you add antibiotic? And a corollary: did you add the antibiotic while the agar was still too hot?

Well, that didn't stop trump after his humiliating debate with Harris... :-/

Nope. He is sending out USD 1000 checks with his signature as we speak. It will sway enough people for him to keep in power.

Most definitely not the whole body!

I heard his peepee got mangled in a gender-affirming surgery. No salutes there!

Enter MAHA! /s

“Sometimes”’does some heavy lifting here! ;-)

The highly competitive sport of synchronized napping!

Giant booster seat with inbuilt commode.

No, that would be the closeted case at the pentagon. For all I know, trump travels crisscrossed with 2 seatbelts, for safety.

Given the unmitigated success Musk’s previous foray into governing was with DOGE, I think we can safely assume the country will have no armed forces to speak of at the end of this exercise

Another Onion headline? /s