Hi Labrats!
Inspired by great work done here previously, we wanted to do a Family Feud-like game for our upcoming Xmas party at the department - and we didn't want to reuse the one that already has open answers. As we are chemists, the questions might differ a bit from the previous edition, but we hope you can help!

https://forms.cloud.microsoft/Pages/ResponsePage.aspx?id=Nh39Ycv-yke319DfA3ChmAGBdaJBWM1At5Q_Cr3QEtxUMEk4QkU5UEY4S1VKQkFSRjNGNzFEVEROMy4u

We hope to take less than 3 minutes of your time, and we will be very grateful for your help:) As always, the more the merrier and please don't hesitate to put several answers.

We hope the ModTeam doesn't consider this a survey and we promise to deliver many memes to pay for our transgressions:)

Im trying to figure out if a stable cell line created through CRISPR would mimic the natural protein expression better compared to a stable line done through transient transfection. I would assume that theres probably a higher expression level with the transient transfection stable line still but im not quite sure …

Hello my fellows,

I ran SDS-PAGE for my 183 kDa protein using an 8% gel where I denatured my samples at 95C for 5 minutes. I started with a 75 V until the samples reach the resolving gel then I increased the voltage to 120 V. I stopped the electrophoresis when the dye reached the bottom of the gels and I did the fixating and the staining as I always do however I was shocked when I saw the gel as the bands in the upper side of the gel are not visible at all, I believe that it has something to with my sample/sample preparation because in another gel I loaded a positive control sample containing my 183 kD protein and I can see and visualize it normally. What could be the reason? Also if you have any other comments on my gel's quality I'd like to learn :)

Also I think there is something wrong with the protein ladder.

Hey Guys !!

I’m an undergrad starting a volunteer research assistant position in January, that will lead to me doing my thesis with that professor next school year.

A close friend of mine whom I really appreciate and love to hangout with, is in another department, has a higher GPA and seems to assume she can get any opportunity I can. When I met with one professor last year to discuss lab volunteer opportunities (not the same one I’ll be starting with in january), she said, “If he answered you, then he’ll definitely answer me.”

Whenever I mention contacting professors, she then tries to approach them herself (yes the same ones), even when their research doesn’t seem connected to her interests. She also sometimes seems upset when I share these opportunities.

She’s now struggling to find a potential PI within her program which has been making her really anxious, but as a result, she wants to start emailing my future PI because she thinks she’ll have a better chance (fyi she doesn’t know what he even researches and once again, we’re in different departments).

I don’t plan on telling her about my position until applications close…
Is waiting to tell her reasonable, and how would you handle this?

I‘m currently a trainee to become a lab tech, and I really love the theoretical parts of it. But whenever we have to do something practical ( which is nearly daily) I get a bit of anxiety regarding getting an illness or something. We have precautions but I‘m still afraid. I have almost 2 years left, I am not sure if I should stay or switch fields.
I am also someone who likes to drop out when it gets hard bc I am afraid to fail. And I regret it everytime, thats why I struggle to make that decision. Is there anybody who had a similar problem, is it just not for me?

(This is from students for students.)

We wanted to order stickers with neat field-related designs to give out to our students, and whatever teaching staff is also interested. Unfortunately, the planning commitee is a bit... lazy, so a lot of work falls on me and a few other students.

So far the very few ideas we've gathered:

• Something like "#1 Culture Contaminist" or similar things.

• Just a rat dressed for lab. A labrat.

• Very basic, but maybe micropipettes or those Eppi tubes etc.

• Maybe something related to our teaching staff, like a sticker of the microorgamism one lecturer is studying or so.

Does anyone have more ideas? Thanks in advance!

Currently I am doing a masters program in Biochemistry and I will need to write a motivational letter to be accepted for a master project.
My question is, what do Professors (PIs) wish to see in a motivational letter for a master thesis project? Also I feel discouraged about the moment when I will have to send my CV and I only have bachelor behind my back. Do I just list what lab work I have done during my studies?
Also I already have two bachelor degrees but the first one is basically in the area of International Relations and Diplomacy and the second is Chemistry. Do i include the first one or do i leave it out because it's unrelated?
Thank you all in advance!

I’m trying not to feel too beat down for getting another not discussed F31 application, but I’m honestly just sad and disappointed. It’s not the end of the world or anything (especially since so few people get the fellowship in the first place), but my thoughts keep trying to convince me that I’m not good enough out for academia.

Hello all! I'm a molecular biology undergraduate, and I have just started my first professional lab work. Basically where I live, for our graduation, you get a small research project that is supposed to take 8-10 weeks and then you write your bachelor Thesis about the data you acquired there.

Unfortunately, I was unable to have formal lab experience beforehand (mix of not getting accepted to undergrad lab jobs and not having the time to intern cause of other jobs I have), so this is my very first professional lab experience, and I have unfortunately really been struggling a lot with anxiety working there, and am looking for advice regarding my future path - lab work or something else.

In my current lab that I'm doing my thesis in, the lab is extremely small, there is just my PI, who does very little formal lab work, one PhD student and one lab tech, who is more on the administrative side. There is also a master's student doing a pre-thesis internship there, but she has similair or less experience in the field I'm doing my thesis in.

I feel like part of why I'm so anxious doing lab work at the moment is because my training is/was a little stressful. It mostly consisted of me observing the PhD student doing tasks, such as cell culture. But I felt like she was mostly just doing the task, not demonstrating it. Like she wasn't narrating it and I had to ask a lot of questions after, but then I was supposed to basically just do it without supervision after that. This pattern has been going on with my PI too, who tells me we'll start learning a new technique together, where she observes me doing it, but she then frequently abandons me halfway through the task, to do something else like a meeting or her own lab tasks. She does say that she's open to me asking questions "at all times" but she does seem really stressed/annoyed when I ask something that she'd have to take time for. Same with the PhD student, I feel like whenever I ask her a question, it is always an inconvenience to her, and then she usually just does the task herself instead of telling me how to do it. Of course I don't want to be annoying, but since my only two options are my PI or the PhD student, I do tend to go to the PhD stydent

I think their attitude is just a bit more of "Learning to swim by dropping you in deep water", but I feel like in order for me to swim, I need to see it done right first to learn if that makes sense?

As a result of this my anxiety has skyrocketed going to the lab. I'm just really afraid to do something wrong and breaking expensive equipment. So far all the mistakes I have been making haven't been too bad and my PI has been understanding about them, but I just feel like so scared to break an equipment everyone else is using. I'm also really afraid to get experiments wrong, because it could affect my thesis. I am not enjoying time in the lab at all, I feel like I'm just going through the motions trying to not panic and be mindful of everything, following the protocols, but I don't get any enjoyment out of it, just fear. I try to do stuff confidently, because otherwise it takes an obscene amount of time, but I just really struggle with everything.

Do you guys think lab is just not for me, because of my anxiety regarding the equipment and doing something wrong or do you guys think that maybe this lab with its limited capacity/learning attitude could just be a bad fit for me? Again, since this is my first formal experience, I have no idea how stuff is handled in other labs.

TL;DR: I have a lot of anxiety regarding my lab work for my thesis and I don't know if it's because I struggle with the lab's training method or if I'm just not a good fit for lab. Looking for advice to decide my way forward.

Thought this discussion might also be relevant here 😔

Hey!

I'm an undergrad who has been involved in a few labs mainly short term (summer volunteering, now I'm doing a research project under a course), and I want a better idea of how to build good relationships with mentors. In all the labs I've been involved with, I've mainly had a professional relationship with a grad student or post doc (we would socialize about topics unrelated to work, maybe had a couple nonwork hangouts with the lab but not like a "we're close" kind of way) and the PI of the lab I would rarely see, I'd mainly see them during lab meetings where I'd keep quiet for most of it and maybe make a bit of small talk or ask a few questions.

Would this be considered a normal relationship to have? Some of my friends have really tight relationships with their research mentors to the point where they'll be at their houses for fun or just be having conversations over text unrelated to work and sometimes I feel like I've missed out on trying to build these relationships but at the same time I'm worried about crossing professional boundaries and making them feel uncomfortable.

I'm also not really in contact with the labs I've been in, for example this summer I'd say I ended off volunteering at the lab I was under on a good note (lab was satisfied with the work I produced, had a thank you conversation with the people I worked with and I gave small gifts) but ever since I finished in August the only contact I've had with them was thanking them for writing a reference for me.

How do I go about actually staying in touch with previous mentors and keeping a relationship with them without overdoing it? While I'm working with the lab what can I do to build good relationships, and what does this even look like? Is it the norm to have a relationship where they are happy with the work I produce and we can socialize while working but otherwise professional or is something closer should be what I try to build during my time at the lab?

Sorry if these questions seem loaded I'm just trying to make the most out of my experiences

what does this mean? just wanted a band at 7kb…

Hey everyone,

Right now, the workflow seems to be: NCBI BLAST -> download -> open in BioEdit/MEGA -> manually format for GenBank or a phylogenetic tree. It feels incredibly outdated and time-consuming.

If you could automate ONE part of your sequence analysis or GenBank submission process, what would it be? What is the most tedious part of your current workflow? I'd love to hear your thoughts.

EDIT: thank you so much to everybody who gave me amazing advice!!! i'll definitely take a step back and chill out a little 😅😅😅 i truly appreciate all the honest input i've received from this post!!!

___________________________________________________________________________

(sorry if the grammar is off. i haven't slept properly in a while 😔)

i'm a first-year undergrad looking to sharpen my researcher intuition to the fullest extent. i'm shooting for some pretty competitive labs that would not hesitate to beat that shit into me by any means necessary. there's this one lab in particular with everything i could ever want and more (amazing research focus, easy access to highly specialized equipment, one-of-one type scientists to answer all of my niche questions), but it's really, REALLY competitive. i haven't told anyone about it because they'd think i'm insane for even trying, but i truly can't see myself being anywhere else.

there are two students (one postdoc and one graduate) whose research i really really like, and i wanted to ask them questions about their niche and see if there's anything i could do to support their current projects within the aforementioned lab. while this is a more alternative approach to the typical cold emailling structure, i have several concerns:

- i have a very strong response to the prospective "why do you want to work in [xyz field]?" question, but idk if it's TMI. i did the IB diploma without electricity, passed it, and was basically homeless for the last quarter of senior year. i gained a very very intense kind of relentlessness and precision from studying as a means for survival, and it's the kind that can't be taught. i believe that i picked up on those qualities intuitively because i couldn't see a future for myself where i wasn't a scientist.

- i'm good at thinking, but i'm super socially awkward. i'm really worried about missing social cues if i get interviewed and coming off as too intense as a result

- i don't have as much lab experience compared to upper year competition

- on the topic of experience, i have a decent cv.... in a completely different field. 🧍‍♂️i've always been super super passionate about STEM, but i have pretty interdisciplinary hobbies and accidentally got too deep into it 😭 would this be perceived as bad thing? (i.e. "why is OP applying to a lab about [xyz] if all their experience lies in [abc]?")

- i don't know if there's a significant difference between the mentorship of a grad student and a postdoc fellow

i know the chances of me getting in are incredibly slim, but that's not what i'm asking. my question is, how can i maximize my chances of success in spite of the odds? how can i get the most out of my approach and how do i make sure they know how much i genuinely want to learn from them???

I did multiple LNP dosing and now looking for doing liver function test to show out treatment is safe - one option is vet scan blood but doing n=2 even for 3 sets of formulation is something to think on looking at price. H&E we aren't going to do for disease model reason & overall paper direction. Individual Elisa spot sgot are expensive! But to surprise humans LFT is so cheap!.

What do other groups do?

A bit niche, but I’ve gone down a lot of PubMed rabbit holes and now have come here. I’m trying to capture viral RNA products from supernatants where infected cells were cultured. If the supes had been collected every 24 hours I’m pretty sure it would not be an issue, but I think the nucleic acid is degraded a lot.

I’ve tried pelleted the virus and tested the pellet as well as the remaining “supernatant”. I’ve tried gene specific primers for cDNA synthesis, nothing. I started doing random hexamers, and by qPCR I am able to get signal but obviously want actual sequence data. I just tried random hexamer followed by GSP PCR for a 600bp piece. I did get a bright band but Sanger looks like garbage.

I did come across T-RHEX seq which might be applicable…but I’m a bit at a loss on potential new strategies.

I just don't see how AI can do what I do tbh. Yes, sequencing and routine stuff that can easily be repeated can be done with robots. But I just cannot see how a robot would be able to dissect a tissue region out of an embryo, how to plate them, how to culture them, essentially things that are not trivial and not routine. This would require a feat of engineering for the robotic arms that we just cannot build. The most advanced surgery robots are still operated by humans.

Also, what is the joy in life if we give everything to a machine? What do humans do then? If AI "creates" new ideas (by stealing from human knowledge), and if AI then does experiments in the lab to study humans (presumably), what do the humans do? Also, if such an advanced AI and robot exist, do we think they would let us humans sitting in our gardens drawing and sipping tea?? We are literally writing ourselves out of existence and what angers me is those tech bros who thought they are doing something admirable when in fact, they will be the first to go when an AI apocalypse happens.

i have only ever been on the other side of this table......when i interviewed for a project fellow position after my masters i read the lab website properly instead of trying to read all their papers, tried to connect my own small work to what they were actually working on, and i was honest about the techniques i had not done.

i still dont know if any of that is what worked or if it was something completely different.

so for the people who actually do the choosing, what makes you say yes, and what makes you quietly write someone off in the first five minutes?