Hello, sorry if this breaks the rules but I figured I would give it a go because I have run out of ideas. I am in need of a cryo embedder but they go for $1K or more new online these days. Anyone know where I can get a 2nd hand one and hopefully less than $500. Thanks!!

Hey folks,

we now had two contaminations (?) in two weeks in our stem cell maintenance dishes.

It took 48 hours after the last passage to appear. We use Geltrex and E8 Medium, no other additives. The medium was not cloudy or discolored, so we are not even sure if it is a biological contamination. Has anyone ever seen anything like that?

Thanks

My pd advisor was a big deal and was requested to write a review article for science about the evolution of a thing he had originally cloned in the 80s. I had done evo-devo in grad school. He talked to me and 2 other pds in the lab about the article and asked us separately to review his draft. I said hey you need to add two paragraphs like this and add a figure like that. He asked me to do some genome searches to figure out how the genes were structured, so I did that and coughed up the numbers for the departmental graphic artist to use in making the figure.

I was not an author, and I also wasn't acknowledged (no one was. He acknowledged the contributions of the entire lab.) I feel like even though I didn't draft text word for word, I did edit the thing and provided concepts that were later included. AND I coughed up numbers for genome alignments.

Recently my local uni offered a 4 month hybrid biotech technician course with the fourth month as a practicum. It's grant funded so I didn't have to pay anything. Cost was always the largest barrier to me going to school, that and being first gen, having hangups about how meaningful a degree is, feeling if I truly cared for something I could self study, general bad experiences with compulsory edu. But I decided to sign up for this course when I read about it with hopes of entering a career with a better growth or at least pay ladder, to do more intellectually stimulating work, for a better chance for my peers to have similar work ethic, and just to experience something new, to go to new places and have new social experiences.

Almost every other labmate of mine has BS is a grad student or has experience in industry, and said they took this course because they couldn't find a job and it has direct placement. I definitely have catching up to do, my labmates say they don't even read the course material but I've been poring over it taking extensive notes, doing extra research and trying to learn as much as possible.

But I was surprised. Most of my labmates seem bored and disinterested, I found myself being the only one to start conversations, answering the most questions, being the most willing to pose new questions, and taking charge of each lab session with my partner. It's probably nothing new to them if they've already taken many classes, but still, I definitely entered this thinking I'd be the one needing to follow and learn from someone else's lead. The professors are pretty hands-off too, the course is very much sink or swim, since day one they have been drilling "lab tech is often also LSO, this is your responsibility" and so on.

I had to return some papers to my professor and he told me to bring them to his office, since I'm non credit I don't have a student badge and had to get some other students to let me in through multiple locked doors in this very serious biotech building. Standing in a service elevator that smelled strongly like animals and had animal-related warnings on it I had a strange feeling of foreboding... not that I have it, but if imposter syndrome were ever to be validated, I am essentially a random guy off the street. Have I really been conducting myself appropriately? Is this initiative I have been taking really the best? Why does this course exist at all? Where is it taking me? Surely there must be a catch, there must be some terrible work that absolutely no one wants to do. And I also feel the perplexing remorse, maybe despite my perhaps illusion of a better career it won't take me anywhere interesting after all, maybe I will end up like my bored and disinterested lab mates.

Sorry for the essay, I just needed to get my thoughts about this out. I am thankful for any kind of feedback or thoughts on the situation.

Many years ago I did a PhD in Cell Biology and then spent 5 years as a cancer research postdoc. My skill set at that time was mainly cell culture, western blots, immunofluorescence, molecular biology techniques etc. I have a decent publication record from that time. I left research for family reasons in 2009 and have had various jobs in science since. Since 2012 I’ve been in a desk job in pharma. However, I’ve been bored stiff for many years, and now my kids are older, I am considering a return to the lab. I don’t want (and probably wouldn’t be a candidate) for a postdoc, but I am interested in getting a technician job and perhaps building up to a senior technician or lab manager. I‘m based in a big city with several universities (not London).

My question is whether this is hopeless. Would I even be considered or are my lab skills completely out of date? Will anyone even consider me? I’m happy to take a significant pay cut and come in at a low band and work for a couple of year to rebuild my skills. However, I’m conscious that I’ll be competing with new graduates with more recent lab experience.

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)

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.

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.

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!

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!

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.

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.

Hey everyone, I’m interested in pursuing the clinical lab science degree at stony brook and will start completing my pre reqs next semester. Can anyone who’s in or has been in this program please list the stats and ec’s that got them in as well as gpa? I’m super nervous since it’s a competitive program.

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.

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

Feel like having fun. Hit me with your best shot.

Hi everyone, I’m a psych 3rd year undergraduate in a psych lab and have been since the end of my 2nd year-so about 6 months now. I’ve loved being an RA and on the side also on the side working on a project in the same lab co-authoring a paper. As an honours undergraduate student this was amazing exposure for me and I’m so grateful i could learn about such an advanced cool statistical analyses. I should mention i was supervised by my lab PI as well as the post doc in my lab (both were co-supervising me, although my pi ended up getting very occupied with other things so he was actually barely involved and would never attend meetings, my post doc would occasionally update him on how the analysis went and how we could make the study better etc, but it was mostly my post doc who guided me through writing the whole paper as well as how to interpret results etc). I was a bit heartbroken once i finished the whole paper and submitted it to my pi, he said since our results are negative and journals wouldn’t want to show our negative results so it may not be a good idea to publish right now. Is this a red flag or does this happen often in academia? I was also hoping my first publication on my cv would look great but now that’s not possible either and idk if it’s appropriate to ask if we can do a pre print. I feel like our findings are still valuable to other researchers and they may waste time doing the same study as ours, it feels like i put in so much work into it and it’s going to be a waste. I can’t help but think my pi just dosent want me to publish or if he really thinks the findings are not good enough to publish even though it’s filling a big research gap. Do i reach out about the pre print?

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?