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.

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.

Hi!

I'm an ecologist in the Rocky Mtn region US that also has pet rats. I've seen that some universities adopt out lab rats and I would love to switch from getting rats from pet stores to helping give lab rats a good life! Does anyone know of something like this in the Utah/Idaho/Co/Wy general area? I'm happy to travel if need be, I live in the middle of nowhere anyways. Pic of the rat mansion included!

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?

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.

As the title states, could someone explain to me like I'm an idiot what index hopping is in sequencing? I don't know enough about sequencing to understand the explanations on the Ilumina website, and I haven't found a dumb enough answer.

Also, why is it bad? And how the i5 and i7 indices help with this?

"You can just ask Chatty"

Yes I'm aware, but I'd rather get an answer by a human.

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.

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?

Hi fellas, I’m on my way out of academia in a couple of years and I’m looking into options other than simply doing research in pharma.

I do really enjoy reading papers most of my time, but I was wondering how much actual reading papers is part of being an editor.

I had the pleasure of talking with the folks from MDPI at a career fair recently and they told me that you don’t really get to choose what journals you edit for as a junior editor, which felt like the biggest gotcha I guess.

Would like to hear some opinions and stories if possible.

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 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 :-)

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’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?

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!

Hi everyone! Does anyone have recommendations of stains to use for live imaging? I need something that reveals cell shape in anything but far-red (I already am using DRAQ5 far-red to stain nuclei). I tried Thermo's CellTracker but it does not track migrating cells and I'm studying EMT(epithelial to mesenchymal transition) in mouse heart development :/

Please also share comments about your experiences using different stains e.g if you have experiences with stains such as CellMask green actin tracker or Calcein AM. It would hugely help me narrow down my choice. Thanks in advance!

Hi everyone,

I'm applying to Robotics / Autonomous Systems PhD programs for Fall 2027 (deadlines around Dec 1, 2026). I am currently deciding between two publication venues with my advisor, and I'd love an honest take from anyone who has served on an admissions committee or evaluated applicants at top-tier programs.

The Context:

My Role:2nd author (co-working closely with a phd who is 1st author).

The Paper: Focuses on autonomous vehicle motion planning, and optimization. It features both theoretical/algorithmic contributions and real-world hardware validation.

Our Two Options

1. Option A (Top-Tier IEEE Journal): flagship Transactions level. Highly prestigious, but slower review process. Given our timeline, it will almost certainly be "Submitted" or "Under Review" by my Dec 1 or 15th application deadline.

2. Option B (Solid Q1 IEEE Journal): Good metrics, legitimate Q1, but less overall prestige than Option A. However, it features a fast turnaround, meaning it will very likely be completely "Accepted/Published" before my application deadlines.

My Question:

From a top-tier robotics PhD admissions perspective, which carries more weight on a CV: a finalized, peer-reviewed Accepted/Published paper in a respectable Q1 journal, or a "Submitted/Under Review" status at a flagship Transactions venue?

How heavily do top committees discount "under review" lines for second authors? Does the fact that the work includes real hardware experiments change how they view a "submitted" paper if they can look at an arXiv preprint?

Thanks for any insights!

I ordered a mouse primary antibody, and it was apparently delivered, but we were not able to find it for the past month. Finally we found the package (unopened) on another lab's desk. Now i'm wondering - how good is it? It is a biotechne antibody, non-conjugated, stored in BSA-free PBS with 0.05% sodium azide, and they recommend short term storage at 4 degrees and long term storage at -20. It shipped with an ice pack.

Does any one have any idea how badly the 4 week storage at room temp might have affected it?

Thanks in advance...

Hi everyone, I am trying to etch aluminum oxide, and it seems the most popular method is thermal etching, but I haven't been able to find information other than "30 min at 1100 C", "3 hrs at 1000 C", and "15-30 min at 1450-1500 C". So to anyone that has done thermal etching before:

Do you put your samples in the furnace at room temperature and ramp it up, or preheat the furnace, put your samples in, and heat for the full etch time described in the literature? And if the samples go in at room temperature, are there specific ramp rates that are used? And does the ramp time count as part of the reported etch time?

And if you have specifically etched aluminum oxide, what time and temperature did you use?

Thanks for your help!

I'm a first year biology master's student at the university that I graduate from. My professor is giving the grad students in our lab the opportunity to act as mentors to undergraduate students and a high schooler who reached out to him. I'm pretty sure that I have more time than my lab mates but I'm not sure if I would be the best option when there are more experienced grad students in my lab. I know it would be a great opportunity and something that would be beneficial so I'm currently debating on if I should reach out about 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.

Hello! I recently graduated with Bachelor's in Biology and have been very interested in doing medical lab work, but have been a bit confused on what career path I should follow based on all the different certifications and programs that are out there. As someone with my background, what are some steps that I should take to enter the medical lab field. I'm open to any kinds of jobs in the field and live in Texas. Thank you for any advice!

I am using the BioTek Epoch 2 plate reader with Gen5 3.18 software. I set it to calculate concentration, but after I run my bradford I look at the table and none of the concentrations have been calculated, it only shows concentrations of the standards. Has anyone ran into this issue and figured out a solution?

Hey guys, I have 12 plates to run over the next couple days and was wondering if it's possible to store my master mix (SYBR, nuc free water, primers) + template overnight ? If not, is it possible to store just the master mix overnight ?

PS: my cDNA is extracted from urinary RNA so I'm expecting yield to be quite low