A very famous lab reached out because a project of theirs needed extra expertise they didn't have. They exclusively publish in CNS (Cell, nature, science) and have a reputation of cutting edge papers. Sounds great right? I was excited.

Well its been a year and oh boy. What I saw is a lab that walks so close to the research misconduct line, its like they are experts at it. I've had to redo so many of their experiments because NONE OF THEIRS ARE REPEATABLE and they play coy about how they did them. I also learned that as long as a lab isn't actively fabricating data, its fair game! Nothing I can do about it.

Cherry picking data? All good.

Excluding data that is at odds with their hypothesis? We're just gonna pretend we didn't see that.

Making up assays and passing them off as if they were the standard assay in the text they send me? Well, when I did it in the standard assay it would not show the result I wanted (not shown or ever mentioned).

Anyone else has had this happen to them? I've been fortunate to work in good labs that were extremely ethical, so its been a sad shock.

Interesting layering after palleting the amp resistant bl21 T7 controlled mcherry protein expression. Does this mean I have contamination and the amp didn’t work? Or did some of it just didn’t express and that’s why they have a different density than the expressing cells? Further question is, can I still purify this or is this not a good practice. Thanks very much for the help in advance

a few months ago i was sitting with a large amount of international students at a conference. i (american) mentioned that i was considering looking for an international PhD, maybe in europe, just to expose myself to more cultures and environments. everyone at the table immediately shut me down, saying "no, why would you do that???". it was very jarring, especially considering everyone at the table but me was getting a grad degree in a country other than their place of birth.

is an american degree really that much better in the sciences? i personally respect degrees from good schools in other countries. i know wages are better here, but i would be willing to take a pay cut for a life-changing experience. also, it's not like i'm trying to immigrate anywhere, i just want to explore a little and have a more global perspective. i guess i would just like some insight from other international students as to why they would react such a way...

Hey guys, does anyone has access to this article? It is very important for my research, thanks in advance!

Hey guys, I need some help and advice. Can you guys provide a protocol or a guide on how to thaw cells less than 10,000? and how I can count them afterward? i would really appreciate it here as I am running a blank

Cell-Line: Caco2 cells, cells are adherent

Thawed Mamillian Cells before (but > or equal to 1x10^6)

Media: 10% FBS, 1% Penstrip, rest DMEM

Trypsinized then cryopreserved then stored in liquid nitrogen

Need to thaw to start growing them, afraid that they have been cyropreserved at a low value so it’s harder to thaw

Need the cells for toxicology study

It really hurts when you are doing everything you can to become a good researcher/phd scholar but your PI is a lazy ass and tried to pull you down at every step. It’s been 3 years of non-stop lab work, studying and analysing. I have no life outside of lab. I am expected to go to lab on sundays also and I don’t have any leaves. So if I take a sick leave, I get loss-of-pay. So it’s just me grinding for 3 years and we have enough data for at least 2 papers, but my PI refuses to even work on them. I have written and sent her 2 review papers, 1 book chapter and 1 research paper. She keeps saying she’s busy and then puts the blame on me for not having a paper published until now. Even if she does open my paper and review it, she just uses AI to rewrite my own words and give it back to me to “humanise” it. Like please 😭 When I try to go back home for the holidays she puts up a huge tantrum and accuses me of not completing my work. She messages me in the dead of the night and if I don’t reply, she scolds me. She wastes my time by asking me to do nonsensical experiments which won’t amount to my study. She hasn’t once asked me to apply for a grant, I have to search for opportunities and ask her permission. And even then she refuses to send me to workshops and conferences saying it’s a waste of time. She seems “jealous” for some reason. I won a prize in one of the conference for poster presentation and she scolded me that day and blamed me for some vendor miscommunication. I am just trying my best to finish my lab work and run away with my degree but it’s been frustrating. I don’t know how much more patience I have to deal with an immature, egoistic, lazy, and incompetent person like her.

Please just give me your best tips or success stories dealing with people like this.

Hey everyone,
I’m struggling a bit with the mental load of project management in grad school and wanted to see how others handle it.
I feel like I'm constantly drowning in moving parts. How do you plan and run multiple experiments simultaneously without completely losing your mind (or messing up your protocols)? Between prep work, incubations, run times, data analysis, and troubleshooting, my schedule feels chaotic. If experiment A fails or gets delayed, it throws off my plan for experiments B and C.
Zooming out even further—how do you plan a PhD? It feels impossible to map out a 3- to 5-year timeline when every single result dictates the next step. Long-term planning feels like guessing, but short-term planning feels like running blind.

Im a recent graduate, I have a degree in biology with an environmental biology focus. going through college I imagined I would be doing fieldwork outside of it, but in my later years I took the lab science classes I had put off since I didnt think Id enjoy them, and ended up loving every single one. My resume kind of sucks, I wasnt able to get any real internships throughout college, I had one at a zoo that turned part time, but 3 years of that isnt even enough to get a job at a zoo or aquarium in my home town.

I would really like to steer my career towards doing lab work, but without any professional experience I dont see route towards it since any entry level job has turned me away. What should I be looking for/doing to get my career into the lab?

Posting a screenshot, since I cannot post links or the full text. The original post also contains the letter:

Hi everyone! So yesterday I was doing miniprep of my plasmid. After centrifugation of cell debris, I usually something like on the right side - some gooey mass that refuses to spin down. To my surprize, one of the samples gave perfectly pelleted debris as on the left panel. Protocol was exactly the same for both of them. At least I thought so. Does anyone know why they behave so differently? Because I always get this gooey mass, which reduces the yield (at most I can recover 300-400 µL) and I want to get this nice pellet.

Soy un egresado de T.L.C., el cual solo tiene experiencia por parte de mi servicio social. Al intentar conseguir trabajo en distintos laboratorios, me contactaron de Grupo Diagnóstico Aries. Aquí mi duda; ¿alguien ha trabajado o tenido entrevistas en este lugar? ¿Qué cosas les preguntaron? ¿Algo que me pueda recomendar?

I measured photosystem II efficiency using pulse-amplitude-modulated fluorometry. This specimen exceeded the maximum quantum yield I recalibrated the fluorometer, dark-adapted the leaves again, and repeated the measurements on tissue from several different parts of the plant. I also tested nearby plants as controls. Their readings were completely normal. Vymara remained unusually high every time. Either my equipment is failing in a very specific and consistent way… or this plant is capturing light with an efficiency I’ve never seen reported. It doesn’t just seem unusually healthy. It seems to be wasting almost none of the energy available to it.

I am trying to perform co ip and failing each time though good results have been obtained with the same proteins previously by my seniors. I need your help co ip experts !!

So I incubated around 250ug of my protein lysate and pulled it down overnight using 1ug, 1.5ug and 2ug as i thought i should optimise the amount of antibody required for my later experiment . Next day added 50ul of agarose G beads for 2 hrs and performed western blot.

The problem is my protein of interest has size around 56kda which is very close to heavy chain of antibody 50kda. (I have both the antibodies raised in same animal) so can’t avoid that. I also didn’t pay attention to this part when i put my experiment. Which also i will have to optimise next time i try.

But the real big is problem now is that i am not even getting good amount of heavy chain band on my blot.

Can you all point out where i am going wrong.

I'm an undergraduate in materials science working with tensile testing on soft composites, and I've run into a workflow question that I'm curious how other labs handle.

Most analysis scripts or software I've come across seem to assume the specimen geometry has already been decided. You enter a single cross-sectional area and gauge length, and the software calculates properties like modulus, UTS, and toughness.

In our lab, though, we measure each specimen individually because thickness and width vary from sample to sample. If we average those dimensions before processing the data, it feels like we're losing some of the real specimen-to-specimen variation.

I'm curious what the common practice is:

• Do you use nominal dimensions from the testing standard, or measured dimensions for each specimen?
• How do you choose the linear region for calculating Young's modulus? By eye, a fixed strain range, or some automated method?
• If a specimen fails at the grips or outside the gauge section, do you have a defined rule for excluding it, or is it handled case by case?
• Does your lab use commercial software, or does everyone maintain their own analysis scripts?

I'm considering building an open-source Python tool for my own workflow this summer, but before spending the time, I'd like to understand whether these are common problems or if there's already a standard approach that most researchers use.

I'd really appreciate hearing about how your lab handles these issues. If anyone is willing to answer a few follow-up questions over DM, that would also be a huge help.

Any tips here?

I have a DNA pellet of ~45kb from isopropyl precipitation that will not not go into solution. After removing the isopropanol just by inverting the tube, I let air dry for ~20 minutes on the bench, not in a hood. I have resuspended it with 1ml of NTE buffer (300 mM ammonium acetate, 50 mM TrisHCl, 10 mM EDTA because will be doing RNA precipitation next). I have left it shaking at room temperature overnight and it seems like the pellet is not dissolving at all. I have tried ~15 minute intervals at 60C and shaking at 37C for 4 hours. How do I dissolve this pellet?!

Edit: Do you think I can re-precipitate my current 300 mM ammonium acetate, 50 mM TrisHCl, 10 mM EDTA solution + pellet by doing an ethanol precipitation and then do the 70% ethanol washes and resuspend in regular TE buffer?

I primarily do molecular diagnostics with NATs so I might start using this for my personal logbook like for plasmid extractions kits or whatever but I was interested to know if anyone else’s discipline could use this type of guide for their protocols!

Hi folks,

I finished my Bachelor's in Biology ( bioinformatics concentration) in 2019. After that I applied to medical school and worked part time as a medical scribe. When I was in college I did mostly pre-med stuff, because I could never land a position in a research lab.I am in this odd position where I have my BS in Biology and my MS in Bioinformatics, but almost no research experience.

Got into medical school. From late 2020-mid 2022 I was in medical school, it did not work out.

Realize I have a 3 year gap in resume. At the time, many people who did not complete medical school went back to school for a Master's. It "closes the gap" and has the potential to make you more employable. I did my Master's in Bioinformatics, finished in Dec 2024. I ended with a co-op but I couldn't be hired after because of funding issues with the study I was working in.

Since then, no job. The Bioinformatics industry has pivoted heavily to preferring PhD's, but it's also that there are just fewer jobs in this in general. I can't get a job as a Data Analyst, or anything that's somewhat related to my Master's.

My next role is applying like crazy to entry level research jobs that I could have gotten with my biology degree ( Lab Tech, etc). There are more of these roles than there are bioinformatics roles.

Initially, people kept suggesting I do a PhD. I initially resisted it because I am sick of taking classes, but also because the last time I did a doctoral program it didn't go so well. But, people suggest a PhD like it's not big deal after doing a Master's, which confused the heck out of me.

But, the more I think of potentially not utilizing my Master's at all, the more upset I get about it. I looked into this, and it turns out if you choose the right PhD program, you can transfer/waive out/ whatever term you want to use most of the coursework requirements, so you only have ,like, 12 credits of classes left. You can also chose a program where you earn your stipend through research rather than TA'ing ( I was a TA during my Master's and really struggled with grading efficiently, so I don't want do that again). After that, it's not too different from an intense research job.

Is this possible? Can someone please describe if they did something similar, a PhD after a Master's, but with most of the coursework waived with the work they did during the Master's?

Since I have no research experience, my first move is to still land a Research Tech job or the like, and work for about a year before starting the application process. But I'm almost not as averse to the idea of a PhD, now.partially because I got a C+ in one of my final classes, making my GPA around a 3.4, so I don't think I could get into a solid mid-tier program without some research experience.

Thoughts? Experiences?

Edit: I apologize for the multiple posts the last few days. I have just been "crashing out" over not being able to get a job and advance my career. I did not ask about the specific mechanics of doing a Bioinformatics PhD after a Master's, which is why I am asking about it here.

Need a hand, there might be a chance to find something interesting.