Hello!

I did an experiment to analyze the expression of miRNAs related to cardiovascular disease using a QIAgen plate (miRCURY LNA miRNA Custom PCR Panels ). I tested two conditions and obtained differently expressed miRNAs. Therefore, I would like to know what analysis I can do to evaluate which pathways are being altered?

Caveats/preface:

1. NIH funded my training (while it was profitable to my PIs to do so)

2. I met the mother of my children in an NIH lab, and these kids are directly a result of my time there

3. The best people I've ever seen doing real work at the bench were at NIH

Here's the opinion that will inevitably downvote me to hell but needs to be said: the NIH leadership and bureaucracy absolutely deserves what is happening to it.

Should NIH be dismantled and privatized? Absolutely not. Should the people in the trenches doing the work have to suffer anxiety about their paychecks? Absolutely not.

Should it be a playground where a club of elitist PIs gatekeep access to our own tax money while keeping us serfs to Pharma and forcing us into serving the publishing cartels for the benefit of their rigged system? No.

People who benefit from intramural rarely want to talk about what extramural study sections have done to biomedical research in the US.

I voted for Kamala. In a red state no less. I have no love for the MAGAts and have gone zero contact with close family members who joined the cult, so don't claim I'm a bot or a Russian asset. I'm a guy who did his work, got sold a bill of goods by the same people crying about being victims now. I'm assuming there are a lot of us on the "losing" side of NIH's poisonous effect on Academia who also are feeling some schadenfreude right now.

Burn it all down. It was already rotten. Just burn it, let's build something new with the ashes.

I've worked with a wonderful PhD student for the past two semesters who taught me a lot about wet bench research. She was a genuinely great mentor and wrote me an outstanding letter of rec, and I am leaving the lab soon and wanted to get her some kind of gift. I know a fair amount about her interests, but am having trouble coming up with gift ideas related to those. Obviously I could get her a gift card but was hoping to do something a little more personalized than that. I thought this might be a good place to ask for advice.

the images in lif file are not great, how can i enhance them for better analysis on lasx

Here we are. In case anyone was wondering how the federal payment system was going to stop allocated funds, this is it.

https://www.nytimes.com/live/2025/02/01/us/trump-tariffs-news/treasury-department-payments-system?smid=url-share

Title says it all. I know lots of folks in this sub work in academia and are in or have gone through grad school so my BSc in Chemistry and working in industry for the last 7 years might be less congruent with the norm here, but instead of lamenting the ever broader ways AI algorithms are integrating into our lives and work places, I have been trying to understand how I can use it in my day to day at work and how to use it as a tool rather than just be a casualty of its incorporation into everything.

Does anyone really use it for anything besides revising drafts of important emails or how to phrase something differently? I was having some trouble diagnosing an issue with an SEC method and was surprised that ChatGPT gave me some good places to check after having looked through some grad level HPLC textbooks and getting nothing from them. But besides the one off uses I’ve struggled to find it to be useful outside of emails for the most part. How do you all use LLMs in your work that you’ve found helpful? Hoping others besides myself find something insightful from this as well so please let it rip if you’ve got anything. Thanks for your time and hope everyone is having a nice weekend.

Hello! I'm biobanking human liquid biopsies (eg blood), for batch analysis by flow cytometry later. I've been using WBC to optimize the biobanking protocol and neutrophils are completely lost after freezing and thawing. Would appreciate any tips.

Our standard freezing protocol is to freeze in 10% DMSO in human serum, ON in Mr. Frosty in -80 and then transfer to liquid nitrogen. I've also tried Bambanker. To thaw, I add cells to thawing media (complete media plus 10% serum, and nuclease), and wash twice with thawing media.

Hey all! I’m freaking out slightly. I made a stupid mistake.

I work with Non-obese diabetic (NOD) mouse ES cells. We make N2B27 and keep it in -80, then when we need it we use it to make 2iLIF.

When I make N2B27 I first make N2 which is

• Apotransferrin
• Insulin
• BSA
• Sodium Selinate
• Putrescine
• Projesterone

And then to create N2B27 I add it to DMEM + Neruobasal + B27 + L-glut + 2ME.

I made twice as much N2 as I needed in order to be able to freeze half and use it next time. However, someone distracted me while I was adding it to the DMEM and I accidentally put all of it.

Will those ingredients kill my cells?

I don’t have enough DMEM and Neurobasal to dilute it and won’t for at least another week. So I have twice as much N2 in my media. Will that kill my cells? Am I screwed? I’m scared to tell my PI because he’s been on my case about my “careless mistakes” but when he forces me to do three experiments in one day and i’m in the lab for 13 hour days then i’m bound to mess up sometimes!!

Hi everyone, first off, power to ya during these times. No where near a reasearcher/ a 'lab rat', but I worked closely in a lab and your job is scary / no joke. Getting in those 384 wells with no mistake/contamination is too much stress for me.

Anywho, family friends Toxicology / PCR testing lab needed a data entry clerk, and they picked me up. My background being in tech, I started building software that helped automate tasks around the lab.

The 2 main features of my software is:
* Automatically creating test templates for our machines, no longer manually creating these tests, which they were super happy about

* Programmatically streamlining sample check in. They where manually checking sample entries, and organizing for the actual testing process. My software automates that by using barcodes, and assigning sort number for each sample.

Small other features like making daily work list to pass inspection and what not.

So far, since I am not Storing, Transporting, or Processing PHI on any of my computers, I believe I am not needed to be HIPPA compliant. I plan of course to be HIPPA compliant once I grow with this software. I grow, they grow, it's why I want to give them a good price as well.

But for these 2 feature sets, which are almost used daily, I want to figure out a number, and I dont want to overcharge much because I know similar software exists (Haven't seen much advertisements for the actual check in process tho)

Being lab workers, how much value would you associate to a software that auto generates your files for you? I am thinking somewhere around $300 a month? Or am I selling myself short here? Or maybe too much? Thanks in advance!

We treat mice with a drug in their drinking water ad lib. Our usual strains of mice drink this plenty and it's all good. Recently tried a different strain of mice for an experimental purpose, and they seem to be drinking a lot less of it.

The drug water is pretty acidic (pH 3), but the other strains had no problem. We also add 5% sugar in all cases.

We make this solution sterile by using autoclaved MilliQ water (Type 1 ultra pure), letting it cool down to temp, adding drug + sugar, and 0.22 um filtering.

Anything we can do to increase drinking rate? Any idea why some strains of mice would have no problem drinking a normal amount but a different one might?

Does anyone have suggestions on equipment/techniques for sub-microliter molecular biology, for scaling very low volume reactions. I have had good luck with the Echo, but curious what other techniques exist.

I can’t really figure out how it works. I’m seeing some crazy expensive printers but I can’t even tell if they will work for what I want.

I know this is not a question that anyone besides my boss could answer, I’m just wanting some communication with other professionals with NIH and NSF funded jobs. Especially any women/BiPoC/“DEI” in the field of academic research. I have never experienced anything like what we are seeing. I just need people to talk to as my boss is also very confused rn.

With the current state of science and research, I am concerned about the future of my job. My lab is funded almost exclusively via NIH grants and with the executive orders to pause federal spending, I don’t know what that means for the security of my job in academic research. Anyone have any advice?

I sent samples for Sanger sequencing to Eurofin via overnight service on Friday evening. I received results on Saturday morning, but the email mentions 'partial sequencing results,' even though all samples were sequenced. The sequence quality is poor, with almost 80% of sequences failing, despite clear amplification on the gel. Could the 'partial results' indicate that sequencing is incomplete, and they will send another email with complete (good quality sequences) results?

hi everyone im looking for research assistantship jobs in CT (psychology/Neuro) if anyone is aware of any leads! I will be graduating in May and I am looking to do research for two years!

If you’ve been paying attention to the news, major scientific governing bodies that fund research, including the USDA, the CDC, and the NIH have shown they are complying with administration - or have been forced to comply.

As with everything else about the first two weeks in office, everything is in line with project 2025, under the guidance of project of 2025 staff - the scrubbing of federally funded research related to climate change, HIV, and gender has been swiftly and effectively implemented - as explicitly planned in the document. I believe the next step, as laid out in the project 2025 document, is to use the control of the levers pf power and the HHS to demolish our model of national research funding as we know it today. It says in the document that it would be up to states to pay for their own research, that the CDC and NIH must be dismantled.

Money is already tight. Research is expensive. If states are forced to pay for their own research, we are in big trouble. I think its time to try for non federal grants and making more intelligent decisions on how we spend our money.

I am just a scientist. I don’t know how politics works - but I can see the writing on the walls. Tell everybody you know, start talking about it. We need to be prepared for the worst.

Friends - there is still more to learn about how this beautiful world works. I believe there are still people we can help. I believe it’s going to get hard. Lean on each other for support. I’ll repeat the kind words of my close friend “There is always a place for what you’re doing. It genuinely matters.“

My company recently transferred over from just paper notes and physical notebooks to iPads. It was supposed to make life easier for keeping experiments updated and not backlogged, by OH MY GOSH it’s a pain in the ass. I’m Teams messaging my supervisor saying “Hey! These 4 entries need to be reviewed or they’re going to be rejected by QA for not being reviewed within the time period!” And nothing gets done.

Plus, since it’s Microsoft based, you do that thing where you put a single period and it shifts everything to the left or right, which is annoying.

We also have to sign these notebooks but the drawn signatures never stay in the same spot and they float all over the page. You know what signatures do stay in the same place? INK ON PAPER.

I’m with the boomers on this one, I wish we could just go back to paper ;-;

In light of the insanity and frustration we’re all experiencing was hoping to add something light hearted and fun.

Okay so everyone (or at least they should) has a late night lab song to get them through. It’s 7 pm you have cells to passage or you’ve extracted DNA and need to do a bead clean up. You’re somewhat delirious and start singing to your cells. What’s yours? Altered lyrics encouraged. Or possibly I’m alone in this and a little insane.

Take me home tonight, I don’t want to let you go until I’ve refreshed your media

https://abcnews.go.com/Politics/federal-workers-told-offer-paid-september-resign-valid/story?id=118317566

Well I'm just a dumdum. I thought "government" meant a more... Nebulous thing... Like it's power, "reach" and inner workings aka bureaucracy.

Or even people with power. But no, as usual I very clearly do not understand.

So now govt = day to day scientists, inspectors, advisors, admin, so forth and so on. Hmm..

But is not inclusive of POTUS, vp, etc. Color me curious

I can see my protein on sds page and there is a thick band. but there is very low concentration according to bradford or 280nm reading. Please help.

Hello fellow LabRats!

I’m a PhD student and recently my laptop is dying (doesn’t boot/ freezes/ gives blue screen error after 20 min of launching) if anyone is capable of help and save me the Coin id appreciate

With that said, I’m looking for a new laptop. I work with Microbiome analysis, so a lot of data, a lot of R in-house analysis, confocal microscopy, all the deal.

Would you have any suggestions of a laptop powerful enough that doesn’t brake the bank?

Scientific salutes,

A stressed PhD student

Might have ADHD

I’m a PhD student in Switzerland, and every time I do cell culture and my PI sees me, he complains about the way I do it. From how I handle things inside the hood, to washing cells, splitting, and even the trypsin protocol—basically everything. It’s gotten to the point where I start making mistakes when he’s around because I get nervous, and now he wants to “train” me to teach me the “proper” way.

The thing is, everything I do, I learned from highly skilled people before joining this lab—people from a highly renowned place in the US, some of whom published first-author papers in Cell, Nature, and Science. I have a lot of experience working with immortalized cell lines, maintaining and amplifying them, doing treatments for differentiation, transfection, etc. Never had any issues, never had a single contamination.

I find it frustrating (and honestly, kind of offensive) that he constantly criticizes my technique. It feels like it diminishes everyone who taught me before I came here. That said, I get that he has his own way of doing things and wants me to follow his method. I’m okay with that—it’s his lab, and I’ll do what he wants. But I want to have a conversation before the training to make it clear that just because I wasn’t taught his way doesn’t mean my way is wrong.

Has anyone else dealt with this? Any advice on how to approach this conversation without coming off as defensive?

"Out beyond ideas of wrongdoing and rightdoing, citation count and citation farms, h-indexes and impact factors, there is a field. I'll meet you there; where grants, and all the aforementioned things do not exist, where research can be done free of these stupid worldly things, where we do not blindly follow the authority that decides all these things. When the soul lies down in that grass, the world is too full to talk about, and we can talk there freely."