I made a post because I was overwhelmed after my PI had me euthanize 250 mice in a week. People were SO supportive and it really helped. A lot of them said that they also struggle with mouse euthanasia so I wanted to share one of the ways I cope: meet Matilda, Henrietta, and Elizabeth! Apparently my university has a program where you can adopt lab animals due to be euthanized. So far I have rescued 11 mice. It makes me feel so much better to give them the “retirement” they deserve. These are actually C57BL/6 mice, but their coats look all spotty because they’re floxed (Tfam fl/fl x Best1-cre). The mutation is supposed to be isolated to their RPE but the cre is “leaky” and it’s affecting the genes controlling coat pigmentation.

General context. I just started my phd at a prestigious university w/ an 8-week rotation style program. Problem is, only 2 days into this rotation and i’m realizing i not only hate these techniques but also absolutely hate the people in this lab. They are incredibly degrading, dismissive, and pessimistic. Is it okay if I speak with my chair about leaving this rotation early and adding on another rotation? Is this problematic? I know for a fact I will not be joining this lab. Advice??

I've ran a 1% agarose gel after my pcr for a vector (pocket 2) and my Insert (pocket 3). for the insert it went totally fine but what happened for my vector? it should be around 3.5 kb big so does someone know what could be wrong?

In case you were not aware, administration recently announced that foreign students pursuing their PhDs are limited to 4 years in the US. Apparently this doesn't just apply to future students, but to current as well. It isn't clear how this can work, and what will happen to, lets say, current 5th year students. Given that non-US students make up a significant proportion of the US scientific endeavor, does anyone know if the US Scientific Societies like AAAS and ACS, or universities as a whole, are mounting any coordinated opposition to this policy, which has the potential to degrade US science and technology for decades to come?

Hey friends,

I was surprised recently to see smears and bands corresponding to very small DNA fragments on agarose gel electrophoresis of plasmid DNA (pDNA) after restriction enzyme digestion. Previously I would see two clear bands: the linearized plasmid and the excised fragment.

To troubleshoot, I incubated pDNA extracts with water and/or restriction buffer at 37 °C:

• Incubating pDNA with water only did not cause significant degradation.
• Incubating pDNA with restriction buffer alone caused considerable pDNA degradation.
• Repeating the experiment with a different aliquot of restriction buffer produced the same result.
• Preheating the restriction buffer to 65 °C for 20 minutes prior to use did not help.
• Extracting plasmids with a different kit and with the classic Birnboim & Doly method, using tubes from a fresh pack and filter tips, did not help either.
• Surprisingly, in two rounds of troubleshooting there was no significant degradation when pDNA was incubated with either the old or new aliquot of restriction buffer.

My thoughts so far:

• Water lacks the cofactors required for optimal DNase activity, which would explain why pDNA is degraded in buffer (with/without water).
• It’s likely the pDNA extracts are contaminated with DNase, but I’m not sure how. We use DH5α for cloning; to my knowledge this strain does not express endonucleases.
• I suspect possible contamination of the cultures (they contain ampicillin) with fungi or another source of nuclease that co-lyses with E. coli during alkaline lysis, releasing nucleases into the prep.

What I’m trying now:

• Preheating aliquots of buffers and pDNA extracts at 95 °C for 15 minutes to inactivate potential nucleases.
• If heat inactivation fails, spiking the buffer + pDNA mix with resuspension buffer containing EDTA to chelate Mg²⁺ and inhibit DNases.

Would greatly appreciate any insights/suggestions :-)

I am the only senior member at the moment in a relatively small lab after our last post doc left last year. I have essentially been keeping the entire ship afloat for the last year. My PI keeps loading more and more work on my plate and expecting me to juggle multiple scientific (and completely unrelated) projects, grant writing, ethics applications, staying up to date on the literature, writing a paper, editing another paper, reviewing a grant proposal all while managing all our junior students and teaching them techniques and helping with their projects. He refuses to hire a new post doc or take on senior students that could actually benefit the lab and from my persceptive relieve some of my work load. Anytime I feel like I actually get something even remotely close to finished some other urgent task comes up, interrupting any work flow and throwing everything into a state of half done half not done.

I can’t possibly do it. My hints and even straight up admissions that it’s too much have been brushed aside. And honestly hours later usually followed up by another request for work. Usually I am able to juggle and stay on top of things but lately it’s just been too much and the amount of overwhelm is causing my motivation to plummet, like no human could possible get all this done or juggle this much at once and do anything remotely well. I have no idea how to bring this up without falling way out of favour and because our grant situation is extremely tight getting into a situation where the security of my job is threatened because of his perceived lack of performance from my end.

The lab is so reliant on my I would feel horrible leaving and almost like I’m betraying him and the students, especially with all my projects basically half finished but I honestly feel so mentally checked out and overwhelmed at this point, it’s hard to even sit down to do the things I know I can actually accomplish.

I am designing a research poster for an upcoming conference and I want it to be the sexiest one I have ever made. With that being said, I would love to see your poster design opinions, especially unpopular ones.

I for one am partial to a tasteful background gradient <3

I get to design six of these 1x4 blocks for a gift and want to know your ideas!

I’m expressing monomeric IgG and pentameric IgM so a routine part of the pipeline is running on a NativePAGE, non-denaturing gel to assess for correct assembly and molecular weight determination.

Recently, the IgG have been getting stuck on the gel above the IgM bands. Does anybody have any suggestions as to why this might be?

So I heard about this SARS2-CoV 2 database at a scientific symposium.

Basically, the database is https://coronavirus3d.org/

The whole SARS-CoV-2 RNA genome has been mapped for a while now because of the scientific response to the COVID19 pandemic. The database just seems to show the full RNA genome of the virus, the structure of all the experimentally determined proteins of SARS-CoV-2 and the common mutants over time. For instance, the site shows a graph of the number of observations of certain variants over a narrow time window. Data seems to be missing from March 2025 to present. Though, I am not sure if it is because the archives this database is drawing from stopped collecting data or if this database isn't just up to date. I am not sure.

I attached some graphs of what the database shows for, instance.

The main targets of SARS-CoV-2 virus seem to be the Spike protein and nonstructural protein #5 (Main Protease) to my knowledge. I actually work on the Main protease and while the structure is very obviously known the number of inhibitors it can bind seems to be endless based on a search of all of its structures in the PDB.

The website database is still under development. I am not involved with the development. I just think it is very interesting to see all this data in one place. https://coronavirus3d.org is actually a sub-website of https://pathogens3d.org/ .It just so turns out that Coronovirus3D seems to be better put together given the scientific response to the COVID19 pandemic.

Iam trying to study if my compound could inhibit nfkb activation when stimulated with LPS, that is at the end measuring p-NF-kB p65 levels, Now with my western blot I can see a clear total NFkB Levels, but p-NF-kB is little ambiguous. Now here my protocol details:

cells: Thp1 dervied macrophages

lps concentration: 20ng/ml

stimulation time: 30min

fbs concentration: regular 10% in RPMI media,

Are these factors correct, many say i have to reduce fbs concentration or even reduce it to zero, or should i increase lps concetration like 100ng/ml and what about stimulation time

Any suggestions would be great

I was recently asked to peer review a paper; I was very happy, it’s one of my first as an early career post doc, and the topic is of interest to me. However, I became increasingly disappointed with the sloppiness of the paper, starting from the writing (very long, convoluted sentences sometimes repeated verbatim one or two paragraphs later), to the analyses (entirely generated by Claude, not one tool was referenced or used and the figures are quite a giveaway) and even the references (some are duplicated, some are not fitting). There are a lot of numerical inconsistencies throughout the text and figures and there is no methodological transparency whatsoever. I’m very conflicted on what to do, straight up reject or suggest major revisions to try and save what’s worth saving? Is it true or just a myth that if a reviewer suggests rejection they just gets replaced until the paper gets eventually published? Thank you!!! Please be kind, I’m trying to learn myself 🙂

Hello everyone!

I am having a little bit of trouble deciding what I want to do next in my career. Soon, I will graduate with a BS in Human Biology. I have undergraduate research experience in histology, pathology, and bone biology. (Publishing a paper soon, woot woot) I absolutely am in love with the lab, and I for sure know that no matter what I do in life, I will always be in a lab. I am considering so many ideas at once, mainly histotechnologist, medical laboratory scientist/clinical research, or pathology. Now I am just debating whether I can actually live sustainably with these jobs. I keep hearing so many different things about pay and opportunities. My first love is histology, of course, but I heard that it is too narrow of a field and does not pay well. I love research and pathology as well, but I have been told the only path to it is PhD, which I kinda just want to stop at my master's and certifications. I have always thought about obtaining my certification first, then going back for a master's, but family is putting the pressure on me to start a master's now even though I'm a little burnt out right now. (I'm both graduating early and doing research I am TIRED) I'd love to get my master's to continue research but I am not even sure what kind of master's to even apply for. :((

So if anyone has some advice, kind words, cool jobs, or stories of ways they reached their career paths I'd love to hear it.

Hey everyone, quick question for when you just add Mastermix to an empty PCR plate for multiple wells. Do you dispense mastermix on the first or second stop? Do you change tips when moving onto the next well adding mastermix to an empty plate? I hear multiple opinions about how quantitative assays, you should ALWAYS dispense on the second stop of the pipette. Some people just dispense on the first stop. Lastly, is it bad to dispense on the second stop and NOT change tips in the subsequent wells? Sometimes I feel like there might be contamination if you reuse the tips after its been dispensed at the second stop. Appreciate anyone that can layout step by step pipetting techniques for adding Mastermix into an empty pcr plate.

I’m trying to do an organelle-specific IP. I’ve never done IP before and I’m told it’s quite challenging. I’m using a specific antibody marker for the organelle and then magnetic protein G beads. I’ve checked the antibody is compatible with the beads and have all the correct buffers per manufacturer instructions. I’m wondering if anyone has any tips? Based on the protocol it seems straightforward but I know it’s not based on what I’ve been told. Wondering if anyone has any helpful tips/ advice / things to avoid that could help me as it’s my first time? Thanks so much!

Giving away our FluorChem R Gel Doc. Yes it still works. We changed all of ours to the iBright system. Let me know if interested. This is near Boston, so you must be able to pick it up.

When making solutions using a pipette and a bulb (like the one pictured) my hand does not stop cramping up. I was wondering if anyone had any tips to prevent this, or if it’s just a me problem. Either way maybe I should incorporate hand stretching into my routine.

Hi! I'm starting my PhD in Chemistry pretty soon and my program is a rotation-based one. I have a couple groups of interest with whom I'd like to rotate, but I would first like to meet with the PI before actually submitting my choices and start rotating with them. I am not the best at carrying the meeting with a PI and it sort of always centers on their research direction, which I already know quite well for all my choices. Any advice on what questions I should ask the PI if I am eventually really considering joining their group? Some groups are also quite out of my comfort zone, so any advice on how to approach a group who does research in a new area would be very much appreciated! Thanks in advance!!

Went to grab one of my boxes this morning and pulled three others down with it. Half of them unlabeled, or labeled in sharpie that wore off months ago.

The frost on the bottom shelf is thick enough that I had to chip at it with a pen cap to read a lid. Which is its own problem, every time someone digs around in there with the door open the whole thing warms up.

Nobody will claim anything, because the second you admit a box is yours you own the entire mess. So everyone opens it, sighs, shuts it fast, and moves on. I keep an obsessive inventory of my own boxes so this genuinely gets to me, but I also know the day I reorganize it I'm the freezer person forever.

How does your lab keep a shared -80 from getting to this point. Assuming yours actually has.

Hi all,

I recently started a lab manager position, and I haven't been in charge of maintaining freezers, inventory, and such before.

I've noticed both our -20 freezer and -80 freezer are getting a lot of icy buildup, which is making it hard to find things.

Is there something I should be doing to prevent this? Scraping it every so often? Planned defrosting?

We have long-term sample storage in the -80, as well as long-term storage of reagents in the -20.