To give a bit of background I posted a few days ago asking for feedback if I should continue working while on paid time off. My institute originally gave me an option of 1. Be laid off or 2. Go on paid time off for 3 months to burn my PTO and keep health benefits.

Many of you suggested I should not work on PTO as it is paid time off and I agree!

Well today my coworker told me that management and HR have viewed my “I will be unavailable and unreachable while on PTO” as a “change in character” and that I “no longer show interest in my projects I manage”

So they had my coworker tell me they are planning to revoke my option to burn my PTO and lay me off instead.

Is this considered retaliation? I’ve truly never been in this situation before and feel like crap.

I also burst into tears for 40 minutes talking to my coworker because they’re now claiming I lack character and I’ve only ever gone above and beyond.

Any feedback is helpful.

Kinda just need another opinion on this happening today. So I went out of town the week before last for a big life event and PTOd the whole week, and the following Monday. I ended up being short 2 hours for the full 8 hours on Monday, so I made up for it by working late Tuesday- Thursday. Tuesday and Wednesday were like 9am-8pm, Thursday was like 9am - 7pm. I reported Tuesday and Wednesday as being 9 hour days to get me to 80. PI said today that this was a complete misrepresentation of my time, and will be reporting me to HR for time card falsification? He says the extra hours were not approved and that I needed to take the missing two hours as unpaid.

this is probably an easy one :)

Settle a debate for me:

I think it’s absolutely unnecessary to do 3 washes with TBST after blocking, somebody else in my lab does the 3 washes

Edit: forgot to mention this is for a WB

hello labrats!

We run cell-based in vitro screens as part of our work, and the most common mistake we see from teams ordering their first screen is jumping straight to cells before the question is defined. So here is the decision logic we walk through - useful whether you outsource or run it in-house.

Biochemical vs cell-based vs in vivo:

Biochemical: fastest, cheapest, cleanest to interpret. But it only tells you about the molecular interaction, not what the compound does inside a living cell.

Cell-based: closer to real physiology. You see integrated functional effects - proliferation, apoptosis, cell-cycle shifts, pathway readouts. Slower and noisier than biochemical, but far more informative per compound.

In vivo: most physiological, highest cost, longest timeline, most variability. The wrong tool for triaging hundreds of compounds.

For early hit-series triage, cell-based is usually the sweet spot: controllable conditions, decent reproducibility, and it batches well.

What actually decides the timeline: the growth rate of the cell line you pick, the dosing scheme, and how many readouts you stack on top (cell cycle, apoptosis by flow, ELISA, Co-IP, reporter assays). That is why "how long does a screen take" has no fixed answer - the panel defines it, not the vendor.

One thing to decide before commissioning anything: what decision will this data change? If the answer is "nothing specific", the screen is not ready to be ordered.

Disclosure: I work at a contract research organization offering these services. General information only.

I recently started a new research tech job in a lab that does a lot of mouse genotyping. For this particular gene (IL17Cre), none of the PCR reactions seem to be working lately (the issue predates my hiring). The protocol has been the same for years and the reagents are fine. The only major changes every week are 1) the person setting up the PCR reaction and 2) the mouse DNA. Something is obviously not working, but we are struggling to pinpoint what exactly. DNA that works for other PCR reactions doesn’t work for this one, and people who successfully run other PCR reactions cannot get this one to work. Increasing the DNA concentration produces a visible band, but the weird M shape and smearing are still there.

I’ve seen smearing, but I’ve never seen a pattern like this before. I have no idea how to troubleshoot this reaction. Has anyone ever seen something like this before? Any ideas what could be causing this or ideas to try to fix it? We’re not able to set up new breeders because we can’t get this genotyping to work which is very frustrating.

i will start......in my lab the PI gave every one of us our own pair of crocs and they never left the lab. you come in, keep your outside shoes in the shoe rack at the entrance and change into your lab crocs, so shoes never needed any cleaning.

i was working with cell lines there and the cell culture was in a separate compartment inside the lab, so we also wore aprons and sanitised a lot before entering that part.

curious to know how different it is in other places

Annual PM today. I’m the only one in our lab who runs it so it feels nice to know my labor is appreciated by someone other than me 😂😭

Hey guys,

I'm stressing myself out a bit about a new part of my PhD project. I need to knock out a gene in the bacteria I work with. I'm a biochemist with a lot of theoretic knowledge but I have pretty little hands-on experience. I've already done some "minor" genetic engineering, like cloning a gene of interest into a plasmid. My PI wants me "recreate" a knockout we already have as training. On paper I know what to do and how it works. Currently Im the only one doing this kind of work here.

The good news is that my bacteria are naturally competent, so transformation should be relativly easy once I have the right plasmid.

How long would the creation of a knockout plasmid take you to build? With my current workload I only have about 2-2.5 days a week for it. Is 6-7.5 days realistic to get the modified plasmid? Idk how long it usually takes to select positive bacteria. Any tips for the individual steps are very welcome!

Simplified protocol:

1. Isolate plasmid from E. coli and amplify the gene of interest from the template

-> no problem here

1. Double digest both with XhoI and XbaI (from NEB; never used these before, only EcoRI and SpeI)

2. Purify both with a PCR purification kit

3. Ligate and transform E. coli

-> until here I troubleshot a lot of the process

1. Inverse PCR of modified plasmid to introduce an AvrII site into the gene of interest

-> This amplifies the whole plasmid, but with only a partial sequence of the gene. The restriction sites are placed so I can insert a resistance gene, which disrupts the gene of interest.

1. Amplify the resistance cassette from another template (with AvrII ends)

2. Digest with AvrII and ligate

(7. Transform bacteria

-> the partial gene sequence allows for double crossover)

Basically the title, I was putting away my stuff after staining my cells but I forgot to put the FACS buffer jar back in the cool room and only noticed now, it would’ve been out for probably a day or two. The buffer just 2% FBS in PBS but is it still ok to use or should I play it safe and just make a new batch? Thanks!

(...) They increasingly amplify human ingenuity and unlock new forms of creativity, empowering Americans to achieve what was previously impossible across science (...)

https://www.whitehouse.gov/presidential-actions/2026/09/inaugurating-the-era-of-super-intelligence/

(...) Dr. Hye Ryeon Jang, Professor of Political Science at Morehouse College, points out that the rebrand stems from Executive Order 14434. However, she simultaneously notes it remains an uncodified presidential initiative rather than officially adopted federal law.

“We also need to make a distinction that it looks like they sort of agreed, but it’s not really a clear agreement on terminology,” Jang explains, analyzing the subtle friction between the White House and Beijing’s foreign ministry (...)

https://maroontigermedia.com/2026/10/president-trump-rebrands-ai-during-summit-with-jinping/

Hi all! I've played around with notion, obsidian, scinote, lab archives, and a few others and had trouble making them work for my purposes. I'm working in plant pathology, so tracking treatment groups, cultures and their photos is my main task. Any recommendations? Do I just need to dig in harder with something I've already tried?

Ok I get its for the movies plot. Shush. But in the movie he made those magic logs that got the train up to 88mph. On a steam engine. Ok so the guy is not just a chemistry ignorant physicist. He knows what hes doing. He built a time machine, out of a Delorean? And he couldn't whip up a half gallon of gas? He goes to the local bar and tries to run the car on 1885 strong hooch?? That plot point upset me before I ever picked up a chemistry textbook. He had a whole Smith shop. He made a big ass refrigerator! He could have cracked some oil and made gas. Right?? My question is how would you handle Doc Browns situation? How would you make gas in 1885? Or i guess maybe just pay the $80. Nevermind just a chemistry rant.

Does having a penicillin allergy prevent you from working with ampicillin or other antibiotics of this class in the lab? Do you work with it with extra precautions?

I teach an undergrad lab and the next lab is E. coli transformation. We use a teaching kit and the protocol says students with penicillin allergy should not participate in this lab. But the LB ampicillin agar plates are prepared by the TAs ahead of the lab. The ampicillin is embedded in the LB agar, and there is no suspension culture (they count the colonies and are done). The lab is done on an open bench, but I don’t see how they can get exposed to the ampicillin unless they eat or inhale the LB agar or touch it and stick their hands in their nose or mouth. I am wondering if it’s necessary to exclude these students from this lab or to allow them to opt out.

Sent my MSc research proposal to my PI yesterday.

This morning he sent me SMS says “Hi… please call me”.

Not knowing what have I done wrong, I panicked and thought maybe he thinks i used too much AI or something. Decided not to call him and to meet him face to face in the lab, maybe seeing my face would make him less aggressive..

He wasn’t there.

I called him, and he said “Hi, I can’t manage to open the word document, could you send me a PDF?”.

God, felt so stupid for panicking.

He never yelled at me or anything.

Hey guys, I'm looking to get a microscope for my own nefarious purposes (looking at fungal spores). What brands should I be looking at? Follow up question, what supplies should I be purchasing? I haven't had to make mounts for a few years now and the lab work I do doesn't involve microscopy.

Thanks

Hello, I'm wondering about a piece of furniture that I've grown up with in my childhood home and living with currently. It is an antique wooden cabinet with 15 cubbies that what I've learned is for microscope storage. It's possibly circa 1900. In old photos I see that we had previously used it as a bookcase, and in more recent decades storing vases and other knick-knacks. Since I've become more aware of just how prevalent asbestos is, I'm wondering if we've unknowingly been exposed to any residual stuff like asbestos or mercury. Being that I've always been disorganized, I can see how easily it could have been adapted in it's past life as a lab fixture and used to store all kinds
of lab related things, and I'm mostly concerned about the possibility of it having had asbestos or
mercury or anything my OCD mind can think of. I need to go through the contents, some to keep and some to sell, and theres an innocent looking layer of dust behind the glass door, but now I'm
afraid to open it. I'm wondering what do the labrats here think? Should I worry?

Thank you!

Anyone have any easy ways to check for non-obvious cell contamination? I sometimes take 100ul of some media and see if it make 1ml of fresh media/LB go cloudy after a day in the shaking incubator. Anyone got any tips other than expensive tests?