Check your local Value Villages y’all! Apparently Fisher makes kitchen accessories now.

I am one of two lab gremlins at a community college. My colleague sent me a whimsical note this morning of a project he left in my prep lab. Apparently, a student in one of the class labs had attempted to measure the volume of a water bottle with the graduated cylinder in a rather unique way. Which description of his did not prepare me for what reality I was walking into.

The water bottle is essentially near the bottom of the graduated cylinder and is air-tightly fit, meaning the student clearly shoved it as hard as they could into the cylinder. Not sure where the instructor was during their massive endeavor to measure the bottle's volume, but nevertheless the instructor left it for us to discover.

So interestingly, I have a cylinder, stuck in a cylinder, and in the interest of not going through requisitions, it's actually imperative the cylinder remains unharmed.

Suggestions?

Edit 2: SOLUTION FOUND. I explain below for posterity.

I took the common suggestion of heating the cylinder and grabbed a scalpel and a bunson burner. I then super heated the scalpel and proceeded to melt through the base of the bottle while slowly slicing through, reheating a bunch of times. I highly recommend doing this in a fume hood because melted plastic smoke..... don't breathe this. Anyway, once I got the base of the bottle sliced off, I then tried crushing it a bit with forceps to see if I can get a grip and pull it out. That didn't work. So I proceeded to drop the cylinder from a great height a couple times from the nearest stairwell, and it dislodged out after the third loud drop.

Happy to say, my graduated cylinder remains unharmed. The other cylinder.............not so much. Thanks for the suggestions all and joining me for this Tuesday morning random problem.

Edit 1: Attempted solutions:

1. I might need to go searching for stabbier stabby things because right now our stabby things are either too short, too blunt, or making the situation worse (the bottle went down a 100mL into the tube).
2. I tried using a couple spatulas and a pair of bigboy forceps. I got the spatulas to slip around then base of the bottle, but I couldn't get the forceps to get a grip without slipping. I will try this again after putting it in a bath and maybe adding some lubricant to the cylinders.
3. I tried warming up the cylinder and taking one suggestion of swinging it like Barry Bonds, but my hands were apparently still too wet and my graduated cylinder went flying out of my hands. And unfortunately it didn't budge the bottle. I'll try Indiana Jones-ing it in the courtyard here and see if physics can help me.

Final Edit: For the future souls wandering into this thread with similar problems, I hope you are literally the only other person this happens to because any more frequency of occurrence of this will start making me sad for whatever society has evolved around you. That said, hopefully the suggestions by many smart people in this thread can help you out of this jam.

I'm in my 7th year of my PhD. I say 6 years and 6 months. No publication at all. Desperately trying to publish something. Made a barely holding together bioinformatics paper and is getting rejected everywhere. Apparently the topic is too niche or doesn't suit any journal. I do not know why I am posting this here. Now I'm trying to publish a protocol in Bioprotocol. *fingers crossed* But that doesn't cut it because I need two Q1 journal article to get the degree. At this point I'm hopeless and I can't write even one sentence for my thesis. Feel like I'm in this longest downfall of life and the end is not near. At this point I can see a mental depression lurking around the corner of my mind.

I used to write, draw and take photos and teach pupils and had a nice little life before PhD

Now it feels all grey. I do not do anything other than paper work and writing and yet nothing to show. my peers have either completed or in the process of submission. Above all, my stipend is over it's been 2 years I am surviving out of goodness of good old friends (who never choose phd and survived in industry).Barely surving with a credit card debt and personal debt. I'm in my early 30s and still sometime i have ask rent money from my retired parents.

Does any of this pays off at the end? or does it ends at least?

If your work depends on grants, you should review the proposed NIG policy changes, and submit comments.

Late Friday night, the NIH posted a proposal for peer
review changes: "Request for Information (RFI) on Proposed Changes to Reporting Outcomes from NIH
Peer Review". This change would not report the impact score of a peer review grant to researchers and the NIH.

Impact scores give not only researchers an idea of how well they performed, but also guidance on how close their grant may be to funding to re-strategize.

If this will affect you in any way. I encourage you to independent read into the proposed policy changes, and submit comments.

Say who you are and why this matters to you, taxpayer, patient, family member of someone affected by NIH-funded research, and explain in your own words why you think grant scores should stay transparent. Real, specific comments carry more weight than a form letter.

The deadline for comments in October 13, 2026.

From - A badly dehydrated 2nd year PhD student

I have been stalking this thread for a while now, and myself drafted and deleted a couple of posts on the topic I struggle with.

This last whole summer I worked in a lab as a full-time undergraduate researcher under one of the post-docs. At the beginning of my job I disclosed with my supervisor that I struggle with physical disability that often times causes flares of chronic pain and leads to cognition issues. In short: pain = trouble focusing.

At the beginning we built a decent rapport, and work went great. After a while I noticed sharp, frustrated reactions to my clarifying questions and overall lack of understanding that I am not on the same level as them. I brushed it off as the curse of knowledge bias. It has gotten worse, and I reached the wall in our communication. I am stressed to go into the lab, scared of asking questions. This leads to stupid mistakes, which in turn can lead to failing the experiments.

I take ownership of my mistakes, and take notes to make myself a better scientist. However, today my supervisor made an extremely unprofessional and out-of-pocket remark that made me question whether I can continue to work with them. This is on top of often suggesting that I do not think. What should I do now, talk to the PI or just leave?

My PIs installed their own cameras (not professionally installed, just stuck up in the corners of the lab) and made remarks to us like why aren’t you all coming into the lab earlier, we can see you on the cameras (they are rarely in the lab themselves). Is this normal? Feels like an invasion of privacy and kind of stressing me out

Please could I get some honest opinions on if my lack of data is insufficient to pass.

My project aim was:

create an armoured car T therapy for a specific disease: subcloning to create constructs, transducing T cells, testing expression then testing the effect of the activated T cell that also expresses this extra protein on the target cell line in vitro model to see any changes in the ‘disease’ markers.

My progress:
subcloning of constructs completed, sequenced and transduced into HEK cells to check basic expression.
Struggling with transduction of T cells, never made it to activation experiments and only an n=1 to show surface marker expression post transduction
Switched to Jurkat cells (immortalized cell line) successful transduction and proof of CAR- dependent activation.
In vitro models of ‘disease’ state did not work at an experimental level when adding Jurkats, no results here

Haven’t proven activation in T cells, no data of combining in a target in vitro model. Only 5 months left- in the case nothing else works how bad would this be data wise, would this be insufficient to pass? Supervisors much more busy and making no real comment on my sufficiency to pass, of course I will speak to them more on this.

I've got a massive backlog when it comes to manuscripts, 2 manuscripts pending as of now - i do have all the scripts, models, figures, notes etc well documented. But writing manuscripts is something I've been lazy about or idk why I've really been avoiding it.

Thing is, another big project will end in another 4 or 5 weeks, just waiting to get some sequencing done for final hypothesis validation. After that I have 3 manuscripts to write. 🥲

I do have 1 co-author on these but that person is just as useless as me when it comes to this writing bit.

How do you guys do it? Any suggestions or general advice to be more productive about manuscripts?

Also, on a sidenote - I've been getting these ads from jenni & one other similar thing which I forgot the name for..., me and my lab don't really believe in these tools. Do people really use these things? (I'm a pure latex person lol)

Hi everyone,

I made a terrible mistake. Recently I submitted my manuscript and it got accepted. During proofs I found that I mixed up another experiments data and provided as a biological replicate data. I didn't notice this mistake earlier as I copied it from my excel but when I was checking it with my notebook I found it.

So basically I repeated the experiment 2 times with 3 technical replicates but in the manuscript i wrote it as 3 independent experiments with 3 technical replicates. This happened to 2 experiments which I did simultaneously. So if we remove the incorrect replicate the result and conclusion does not change.

I already pointed out this mistake with my PI. She later on checked all the raw data. Apart from this there no other mistakes. She already contacted the journal and they said please take your time to complete proof corrections.

This is my first time and I am spiralling and feeling like an utter failure. What should I do?

Hi, I have just started this new job and there had been no specific mention of how long they wanted me to be there. I am a recent college graduate and decided to get a job instead of going straight into a grad program. However ultimately, I would like to go to grad school and get a PhD. My appointment is one year, which I can renew. Therefore, my plan was to do this job for at least a year and start applying to grad programs, even if I decided to delay starting the program till Winter of 2027.

However, I talked to the lab tech who was in my position and leaving and she mentioned that she would be training my till early December (then she leaves) and I was thinking that I would probably be the next person training the next lab tech. She also mentioned they hired a lab tech before that left after a year and she stayed because she and the lab manager worked really hard to run and create this lab (it is legit a two person lab, run by a single person). She is therefore trying to find a good replacement for her. I am feeling worried that maybe they anticipated I would be here for longer than I was actually prepared for, so I’m not sure what I should do with this information.

Does anyone have any advice on how I should proceed?

So in our lab, we have this giant autoclave. For soem weird reason, it should only take 1-1.5 for entire loading-unloading process but ours took 5 hrs to cool down and depressurize. Did we use a lot of water? Did we load a lot?(We filled two baskets). What to do next time?

Hello, I do not have much experience running gel and I have been trying to optimize on a ~20cm electrode distance gel casting device. This is my second time running the gel and would like some advice. It looks sort of slanted??? at least more than I expected on the first few wells on the very left. If you have any tips Id love to know. I provided the photo below.

I have been checking the gel every 15-20minutes and determined that 1hr and 45 minutes may be good enough to see the whole ladder, though based on my photo I might run it longer at 2+hrs to get better seperation on the larger bp ladders

Specifications:

6x Tri DNA dye at 1:10 ratio.

I ran the gel for about 120V for about 1hr and 45 minutes.

For the buffer, I am using 100mL of 1x TAE buffer + 1% agarose (1gram). For the Ladder, I am using 1kb plus lab ready Thermo ladder. For the dye, I am using 10000x SYBR Safe diluted to 1x. I use of 10uL of SYBER safe for pre-staining.

The liquid and the gel is not overheating. I load about 10uL of the lab ready Thermo Ladder and the same volume for all my DNA samples.

Thanks in advance.

Despite absolutely dumping all over me and my ideas for 2.5 years, now that I have left so she can hire “more competent” scientists (her words when I told her I got into grad school), she’s called me every day for the past three weeks. Texts upon texts over not being able to do the simplest of assays (the ones she claimed I was fucking up because I “didn’t care,” when really it was just because the results didn’t align with the hypothesis. She literally gave up doing an assay that I did every day, because it was “too hard,” then went “idk how you did it all the time geez lol”). I’ve been traveling and she’s been calling non stop. I’ve silenced her notifs but every day I open it and it’s a stream of questions. I’ve delegated everything to the grad students and techs, and told her this, so it feels like she’s just pulling a power move and seeing what she can get out of me for free or just trying to be passive aggressive.

If it wasn’t for the fact that she’s close friends with my new PI, i would have blocked her. How do I stop this? Do I tell on her to her lab manager? Do I contact her boss? Will this destroy my career bc academia is just high school part 2?

Hi,

For anyone who won a postdoctoral fellowship from the ADA and is receiving their postdoc salary from that, do you get the entire amount that ADA sends your institute or does your institution pays you salary at the NIH level and keeps the rest for fringe? ADA fellowship is pays more than the NIH minimum and according to ADA’s policies the institute is not supposed to keep any amount of the salary for fringe. But I have heard from a few of the fellows that is happening at their universities. Is this normal?

TIA!

Hi all,
do you have protocols or electroporating parameters to successfully deliver two different plasmids (8.5 kb each) in HepG2 cells?

The parameters that I selected as optimal when trying with a simple GFP construct seem not to work with the two plasmids, but the overall amount of DNA is the same. Maybe they are too big?
I’m using the neon xt from thermo

Thanks!

EOL-1 cells in RPMI1640 w/ 2mM L-Glutamine supplemented with 10% FBS and 1% P/S

Hey all! I’m hoping to get some inspiration on decorating my bench space to make it more cozy. I work in a lab where students do not have any office space so we are assigned a bench space instead. It’s an ergonomic nightmare but oh well. What kind of things have you added to your space to make it feel more like yours? I should note it is a BSL2 lab which does limit my choices a bit.

I eluted as mentioned in manufacturer's protocol, still this is the 2nd time without any proper bands. I am doing protein purification for the first time. Also, the gel broke (more bad luck).

In the paper from Silvana Konermann and Feng Zhang about dCas9-SAM for CRISPR activation (Konermann S et al. Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature, 2015 Jan 29;517(7536):583-8), a list of guide RNAs for each gene is provided.

Just curious here - has anyone tried any of the guides from the list to see if they work? I know they were computationally derived... I want to purchase a construct with a guide from this list and dCa9-SAM, but it is not cheap, so I'm wondering what people's experience has been in general with using guides from this list. Any insight would be most appreciated :)

Cheers!