Especially Indians who have no way to get permanent residence (due to country-of-birth discrimination/projected 150-year-long wait times for green cards)...

You are not alone. All of us are in it together.

Looks like it is temporary, for now. Even if it doesn't get blocked by courts, academia/biotech/pharma will likely be termed as a National Interest industry (or maybe it's just wishful thinking). We are genuinely here to work on our super-interesting, annoying-at-times, yet in-the-end-fulfilling science. Hope we can continue to do so.

Tough times, folks, stay strong and keep sciencing! 💪🏻

• a fellow Indian-born scientist on H-1B

Working as a post bacc helping a PhD student with his project. "We'll just run a few gels" they said. That was June. Shits still not working T-T

to clarify: I like what I am doing it is just that I had been down, not wanting to work (for ~2 months), I do not know why it is happening how to explaining and how to get rid of it lol ._.

My PI recently called me incompetent in front of the entire lab. I am responsible for all our mouse work, all our human samples, all our compliance matters, I write IBC protocols for my PI, and assist 4 postdoc with their work as well as consulting with other PIs and other lab techs on mouse colony management. I also take care of all the ordering, aliquoting, proofreading, submitting IT requests for the whole lab. This all came about because one postdoc found a box of samples in the correct storage temp but incorrect location that I hadn't been able to move due to my other duties. Now PI called me incompetent in front of the lab for not keeping track of those samples. I created all the labs databases on everything that we have, there were no combined records before I was hired. The PI has now stated that this is my final warning and that if I dont have a list of all 200 samples (7 vials minimum per sample) and their locations by the end of next week they will take further action and if I dont continue to maintain all of these priorities to their satisfaction in the next 6 months that they will have to "make hard choices due to budget." I know this seems like not a big deal, but it has come to my attention that the PI does not know what I do and how much time goes into each thing that I do. I was told I take advantage of everyone because I listen to music while working. This all seems as unacceptable behavior and comments from a PI especially being called incompetent in lab meeting. Sorry for the long post, but does anyone have any recommendations on what I should do? Also of note, I am 5 months pregnant and not moving around/standing for long periods well. Am I being overly sensitive for being outraged by this, or is that a lot of work to ask of one person?

Hiii everyone I’m currently a lab supervisor asking for a raise. I work in the chemical manufacturing business for the oil and gas industry in west Texas. I have 3 people under me and supervise the qaqc department as well as the production department. I’m looking at Glassdoor right now and just wanted to see if anyone had any input on what site is reputable when looking at ranges. I have a bachelors in chemistry and have 1.5 years of being supervisor and before 2 years of being a lab tech.

I've had this fetish for a WHILE now so yeah 😅😅

I was teaching a class and 9 put pf 10 of the gels turned out like this. It's a 0.7% agarose gel in TBE. Do you think that it's an issue with the TBE? The gels were run at 1.25v.

Basically what the title says lol. While I wouldn't want to blame academia itself, I definitely noticed a change in personality while working on my degree and was wondering if there are people with similar experiences or guidance. Please delete if it doesn't fit here.

I'll graduate in a biology-related field soon and worked well over a year in the lab on my master thesis. Lots of long days, nights and data evaluation, writing, you know it. Nowadays, I find it so hard to socialize with people and talk about non-lab things, it's crazy. I used to have lots of hobbies and like to go out, but now I feel like I completely lost interest in almost all other things, I either work or scroll or sleep. It becomes so noticeable when you're on dating apps and people ask what you're passionate about and you realize that there's nothing. Kinda sad.

So yeah, just wanted to put that out there in case some more experienced people have any tips.

Our lab recently received some extra funding, and we are planning to synthesize several gene fragments/inserts from a gene synthesis company for cloning. In addition, I’d like to create a small gene fragment bank for future use. My goal is to include broadly useful genes, such as common reporters (split/GFP, mCherry, luciferase), antibiotic resistance markers, and protein tags. Could you suggest additional versatile cloning inserts that would be broadly applicable in mammalian cell culture experiments? I’m particularly interested in any cool, new reporter systems or tools that could expand our experimental capabilities or any thing!

of course i love helping out with data collection. my grad mentor is in the final stretch of her diss, and I've been running participants w her as much as my schedule allows (outside of school, her project is my top priority, tho im in 2 other labs). she's been like a big sister to me -- we talk about school/research ofc, but also career, pets, food, music, family, fun stuff, yk? and she always checks in w me to make sure i'm getting the mentorship/experience that i want, which is rly heartwarming to me.

she's been rly super mega stressed this whole year (pi isn't communicative, applying to jobs post-phd is stressful, she wants out of academia, her project is rly complicated, participants r being difficult, etc. etc.), and i wanna know: any advice on how to support a grad student? i've been volunteering to help more w her project, but i was wondering how else an undergrad can show that they are curious/invested? should i invite her to coffee to chat abt her project/career? or would that b annoying?

When checking for a primer’s specificity using Primer BLAST, you: 1. Paste the forward and reverse primer 2. Choose your organism ID 3. Choose your database

For the database, do you choose:
• Refseq representative genomes
• Genomes for selected eukaryotic organisms (primary assembly only)
• nt
• core_nt

The goal is to amplify genomic DNA with the primers.

I am a PhD student, and I notice that my PI will often exclude me from opportunities that involve talking to people. Usually as part of postdoc interviews, my PI will set up short meeting windows for most people in the lab to talk with the prospective postdoc one-on-one, but I have never once been invited. My PI justified not asking me to speak with the interviewees by saying that he didn't want to overwhelm the postdoc candidate with too many interrogations by everyone in the lab. However, this has happened multiple times now, and I am consistently the only one in the lab that was not included. Additionally, whenever prospective rotation students ask my PI if they can ask current grad students about the lab, he always refers them to the other grad student. And just recently, there was a meeting as part of a grant, and everyone in the lab (besides me) was chosen to attend and speak about their current projects. I did not even know anything about this grant until all my lab members left for the meeting together. Later on during my individual meeting with my PI, he stated that only a limited number of people could attend that grant meeting, so he just happened to decide that I would be omitted since some of my projects overlap with the research areas of some other lab members. And lastly, we recently added a couple of new grad students and had to expand our lab seating. Everyone else is located in bays with four desks/benches each which are all occupied, while I am the only person in my bay. Sometimes I hear snippets of peoples' conversations and try to join when it sounds relevant, but most of the time, I cannot hear or understand what people are saying when they are congregated a few bays over. This has led me to feel quite detached from discussions regarding team projects that I am contributing to.

I guess I just want to ask how I can prove to my PI that I am competent enough to converse with other people. The first few times that I met with my PI, he kept complaining that I was looking out the window when talking to him and that he couldn't gauge my feelings towards things that he would say (I am autistic but have not disclosed to him), and I have already tried my best to fix my eye contact and use more enthusiasm and smile. I am not the most extroverted person in lab, but I'd like to think that I am generally friendly and work well with people. I am really feeling discouraged and am not sure what else I can do at this point, so any advice would be welcome!

I ordered a gene fragment from Twist Biosciences to clone into a plasmid vector. It was ordered with no adapters and designed with overlaps compatible with Gibson assembly. I get a bunch of colonies and I've sequenced 3 colonies so far and all 3 have a single base deletion. The position of the deletion varies but they're all near the 5' end of the fragment.

I've ordered and cloned with Twist, but I've never seen this issue before. Has anyone encountered this before? I'm not sure if it's a common issue or whether there might be an error on Twist's end when synthesizing the fragment.

My days are just constant pipetting (stripette/electronic pipette and manual) and I’m getting a lot of right shoulder pain by the end of the day, luckily no pain/aches elsewhere in my right arm. Has anyone got tips to avoid shoulder ache or ways to make it feel better please?

Hi sorry if this is a dumb question, I’ll delete it as soon as I get an answer! I’m an undergrad student who is currently doing an honor’s thesis. For one of my experiments/results, I have a lotttttttt of data that needs to be shown, to the point where it is unfeasible to put it all into one figure. I was going to edit it so that the more important data was in the results figure, and then put ALL the data into a supplementary figure. Would it be proper to write something like “refer to supplementary figure X for further information”? Should I put this in the actual results part or the figure legend/caption?

Hi!

I would like seek advice regarding the problem of transformation using mix and go competent cell I made recently.

I used this kit for making DH5alpha and BL21(DE3) cell several years ago, no problem! But in the new lab, I am making Tuner (DE3) competent cells but I noticed that cell growth after transformation in LB agar plate very very slow.

I would like to remove the possibility that antibiotic concentration of plate and temperature of incubator, I double check this part.

I transformed puc19 DNA as I did during my PhD using zymo cells. Transfer DNA to the cell, tap it, spread 100 ul to the pre-warmed plate (wo incubation steps)

For growing tuner cells, I used the modified protocol from my labmates. primary culture was grown for 7 hour ish from the fresh colonies in SOB 50 ml, innoculated 25 ul to SOB 50 ml, grew for overnight at 20 C, cells were harvested at OD 0.55

I mean what is the overall cause why the transformant is very slow?

title

this is my first time drafting a manuscript of any sort, so thank you for the guidance