r/labrats

i want to preface this by saying that i love my job and its my first full time job so i might just not be used to this (started full time a month ago).

i do research at a university affiliated lab and my PI prepared 50 384-well plates for me and my coworker to transfer the supernatent from and add a few more reagents. so we were dealing with 100 plates total.

we only had one multichannel pipette which we needed to pipette anything in or out of the wells. after swapping back and forth taking turns for the first 15 plates (which took us 3.5 hours), me and my coworker realized that i was way faster at pipetting, so i did all the remaining 35 plates while she was helping me out by pouring time sensitive reagents into reservoirs and sealing my plates ect. we ended up finishing at 8:45 pm, meaning i was at work for around 12.5 hours

im a girl and i was really tired yesterday because i got my period at work and didn't have any medication (i cramp really bad). i also didn't exactly have breakfast that morning because i overslept and i didn't know i would be at work for that long because i am unfamiliar with the protocol and my PI didn't give us any warning despite her knowing we only had one multichannel pipette, so i had one meal between the time i woke up at 9 pm when i got home. i was so tired, hungry, and exhausted by the end of the process that i started crying a little while preparing the plates.

my PI is out of town and she sent us an email asking how it went and i responded to her by saying that the plates are ready but we were still at the lab and it was an incredibly difficult process for us to do with just one pipette. she responded with "Great! Don't forget to lock all the doors before you leave." That kind of really disturbed me and I cried on the drive home because i miss my dad and my friends and my ex partner who i broke up with 3 weeks ago (i moved to a new city for this job so no one is around), and my plans to have a nice calm Friday after a busy week were all gone. i was also worried about my kitten at home.

thank u for listening

I think this is more of a vent than anything, but just wanted to get some heavy feelings off my chest.

This was my first real job out of undergrad, and I was really passionate for my work. I was informed with half a days notice that our project is getting cut, and as a result they would have to let me go, essentially effective immediately. I live in an at-will employment state, so that's not uncommon, but it still just sucks.

I'm feeling a deep sense of loss from this, sad that I'll never get to do the exact same kind of work again, and the likelihood of me working with anyone from my team again is extremely low. I keep catching myself thinking about next steps and "to-do's" for my experiments, and have to remind myself that's it's not my responsibility anymore.

Management was really nice about the whole thing, circumstances aside. They offered me rec letters and connections, and made it clear it wasn't about my performance, but I'm still just torn up about the whole thing.

Maybe speaking with a therapist would help me move on? Has anyone experienced anything similar and figured out ways to cope?

Trying not to dox myself here, nor do I need to troubleshoot, but we're working on publishing an in-demand technique that requires using loooong DNA for verification, and I wasn't up to buying all the stuff to purify superlong DNA.

If you think you can provide a prep of 10-50 ug of ~150kb (+/- 50kb) linear DNA at around 50-200ng/uL, we could totally use you on this paper! Maybe someone who works with BACs, etc? Because it needs to be linear, you'd have to have a unique cut site (I think some BACs come with homing endonuclease sites, or maybe some have unique RE's?).

The prep would have to be made with kid gloves: no vortexing, no freezing, only gentle pipetting with wide-bore or cut tips, no sidelong glances, and generally no bad energy in the DNA-breaking spectra. Super duper bonus if you could show us a PFGE run with a pure band!

Any takers? Pub should be fairly high reach! Please DM!

hello, i already did my primer efficiency and will be performing my final qpcr run soon. I need help in regard with the data I need to show in my R&D. I want to know which genes are jn my sample and how much they’re expressed. What data visualizations, analysis and discussions should I be able to provide? Just to make sure… Thank u :) appreciate ur help

here’s what i’m gonna put but maybe i missed some. pls suggest:

• amplification/melt curves by software
• ddCt or 2-ddCt (not sure which one)
• linearity
• box plots

Hey there!

I recently transitioned from an MSc to a PhD in the same lab, focusing on human telomere epigenetics. I’m currently the only student in a traditionally wet-lab environment, which has given me the unique (and slightly terrifying) opportunity to act as our computational lead.

Over the last two years, I’ve established our lab's first bioinformatics pipelines for WGBS and telomere Nanopore-seq (specifically, looking at telomere and subtelomere length, sequence and methylation). I’ve been heavily using AI (Claude/Gemini) for help with coding and pipeline architecture (R, bash, learning how to use my university's servers, etc).

I’m interested in genetics and gene therapy, but also in bioinformatics, and potentially interweaving the two. I want to ensure my PhD years aren't spent just validating my CRISPR colonies and growing cells- but are actually helping me build myself as someone fluent in both the wet-lab and dry-lab/bioinformatics/data technologies that, from what I can tell, are in demand in industry. I think that people at my level, who know how to leverage AI/LLMs, but don't really understand the nitty gritty of it, will not have much of a chance in the data world.

So, some questions for folks in genetics, genomics, bioinformatics:

1. Is "wet/dry lab hybrid" a strong career path? Or are we moving toward hyper-specialization in certain areas?
2. What are the skills you would put an emphasis on learning as a solo student, self-taught kinda-sorta-bioinformatician, and how would you go about learning them?
3. What company names should I be on the lookout for in these areas?

Thank you for reading!

25F from Italy

I have the possibility to leave to go to Netherlands and work for Lonza as a Biotech Associate.

Anyone who has worked there has some insights that would like to share?

How is the work/life balance with the shifts, especially the night ones?

Does anyone have any real world experience with the reliability of the integra assist plus pipetting robots?

As I tend to see them semi frequently on used lab equipment websites so am wondering if it's a reliability issue, throughput issue or something else.

Just looking at alternatives as our large liquid handler is rather temperamental at times and hindering progress.

thanks

I'm planning an experiment to insert a T2A-eGFP fragment slightly upstream of a stop promoter in a cell line (based on the method in DOI: 10.1016/j.stemcr.2024.03.005). Basically, I have to insert the T2A-eGFP fragment between two homology arms immediately flanking the stop codon in the gene of interest, insert that construct into the cells of interest, and cut immediately upstream the stop codon using Cas9. My question - how do I decide how long to make the homology arms?

Edit - i misspelled HDR in the title sorry!

Frustrated on my undergrad PI who hasn't worked on my manuscript to get it published in 5 years at this point. In my opinion this study is little too good to stay unpublished for years and can easily get me some citations. What should be my steps from here to get it published?

thought id give an update on my toxic lab situation (that ive posted here before) and just general motivation for myself and others in a similar situation. mostly because yall have been so kind commenting on my posts thank you. TLDR: mdphd student was getting forced back to medical school early without easy chance of defending before then.

So, my PI actually fought for me to stay a fourth year (pretty easily for her) without acknowledging shes the one who wanted me out of her lab early to begin with (she said it was my committees fault solely). and 2025 went well. she thought I was working hard and such. I got a paper in pre print.

this year begins and overall feel like im doing well. I do a big RNA experiment that sadly needed to be redone. it was obvious some samples were mislabeled (either by me or the pellets I processed in the freezer had been mislabeled hard to say). a big costly mistake, of which I acknowledge and apologize for and begin working to get new cell pellets fresh for the mistake samples, plus a requirement I do extra QC to make sure its all good. for context we can tell which samples would be which by a couple of genes (not too many details because I fear if they'd read this tbh) and those genes can be checked by PCR easily. its taking me time and im busy writing a grant application among other things. And things do slip for me a bit in the sense my physical health issues really affect my energy and hours plus my grant being finished last minute as I busy with health and benchwork and fixing this RNA plus a few things. and I just get plopped with a ton of new experiments I have to do and im overwhelmed. plus class requirements.

Anyway, so I get the conversation once again that im lazy and unproductive and theres no excuse for it. and apparently its believed i have done nothing for three months. and then I do inform my PI about my health and my overwhelm and ask for advice. she seems okay with it. then she degrades me for 45 minutes in lab meeting and implies that im lazy, just doing it for the CV, need people to "hold my hand and do things for me", will cost the lab its existence and imply her failed grant applications may be due to lack of productivity. she also had implied I have issues asking for help maybe its "my ego" (I actually do ask for help to usually no help, or im too scared because shes mad). She lies to the lab thst my grant didnt get sent to the NIH (it did) and that im behind on school work because an email shes cc'd in (i am not. its a task i must do but there was no exact deadline). this time its worse because to varying degrees my lab mates kind of agree with her. person A and her later go on to imply I have an "iffy track record with mistakes and its just who I am as a person." A acknowledges ive made 3 total major mistakes in three years and that does in fact make me an iffy person and its because i work "too fast" or "distracted" (whereas A says he never makes mistakes). ive begun working 12-14 hour days instead of 8. I feel stupid, lazy, like I dont deserve to have ever gotten here. im worried once again I wont be allowed to defend and will have been a failure. It's always like this. its a lab i cannot ever slow down or need help to get back up. mistakes are punishable and a waste of money and time for data. as she tells me there will be 0 tolerance going forward for any mistakes and i am not allowed to make them. she laughs at me when I tell her I am getting to lab 3 hours earlier in the morning. I dont get advice from the lab because most of its just her telling me I need to do every task all at once 24/7 and just "work harder". she once again threatens to make someone else do my project. that id be at fault if the lab shuts down.

this is a lab the school never let me switch out of. they like my PI and disagree with her mentorship style. but say she wont ever fail me because thats illogical even though she talks to influential PIs and department heads to tell them im lazy all the time. im afraid she wont write me a good letter even if I do somehow make it. shes told Person B that she'll purposefully write a negative rec letter to Person C ruining her chances at a phd.

but I had a revelation recently that no matter what even if this woman costs me my dreams of science due to systemic reasons I cant control, I will be allowed back and make it through medical school where I excel. and I remind myself constantly that even if it was true that I was lazy and stupid and egotistical and unmotivated 24/7, none of these ways of communicating it are okay. i deserve mentorship if I really need it, not degradation. I deserve to be held accountable but given tools to move forward from mistakes and not have them weaponized against me as "making me an iffy person" and then told everyone else is perfect and that im the only person who "needs reminding to work hard". I do work hard. and I have faith i will get out of here somehow.

I think a lot about what itll look like. the grief i might have if im not allowed to be a scientist because of her. Or, I do make it out and what i want to do but have to navigate doing it without that toxicity anymore. what kind of trauma will I carry? I really dream about the day I will be able to post here i made it out. And I think my best advice out there is to get a support network that isnt your lab or your PI or the school admin if they wont help or partially agree with your situation. my friends and parents ( +therapist) are the reason I know i dont deserve this. and pray me and you others make it out too. I also am trying to write a prose poetry essay about my experiences here and maybe ill publish it one day. and I also hope I get the choice to be a scientist and mentor people differently, without degradation.

thanks lab rats for always listening and such.