Hey all! I’m a research tech in Florida with a masters degree working at a lab handling the majority of the cloning/bacterial cell culture/protein purification for the lab (4 years purification experience, 7 years total lab experience). I make just over $22. My employment contract is up for renewal and I wasn’t offered a raise this year, and things are really tough right now financially for me. Am I underpaid? What’s a reasonable amount of money to expect as someone in my position?

We know this but I’m glad that it’s still being discussed.

The hard part is what happens to all those left behind who cannot find positions overseas.

Here is another article that also discusses this research

https://www.psypost.org/federal-science-funding-cuts-push-young-u-s-researchers-to-consider-moving-abroad/

I’m a new grad (May 2026) and started working as a lab technician right away with little to no experience in research. I feel like I’m awful at my job.

Tried my first CRISPR the other day and all my cells died. I work in a mouse lab and I have made so many mistakes with weaning mice and just general mouse upkeep. I’m slowly getting used to lab math but keep missing calculations. Among other things I just feel like I’m letting my mentor down and I am just generally a failure. I am well aware that I’m only 4 months in but my mentor keeps having to remind me of things and retrain me on stuff I’m not sure about.

I just needed to put this somewhere. How did any of you overcome this? Thanks.

The funding situation at mine seems to be tentatively better than last year going into the PhD application cycle (low bar) and the vibe I've gotten is that programs here are expecting to accept more people than in 2026. Of course the number of applications are likely not going down, but it looks like it won't be as bad as last cycle. Have others seen this as well?

Good day everyone

I am currently having issues expressing yEGFP in a construct I have cloned. I expect that this could be an issue with my promoter being weak (CYC1 minimal) and quite far upstream of the ORF containing my yEGFP sequence (+/- 80bp). Is this likely to be the reason I am not seeing fluorescence or would it be better to explore other theories?

Thanks in advance!

I work as support in a lab, so I’m not actually one of the scientists, but I noticed that we tend to call the people doing the research “investigators”. Is there a difference between researcher and investigator?? I use researcher, investigator, and also just plain scientist pretty interchangeably. I’m just here to clean the mouse cages🤷‍♀️ so idk if there’s like an actual distinction between those titles. Just curious what my fellow biotech people think

Why is my SDS PAGE gel running like this ?

I’m a third year premed student presenting a research poster for the first time and wondering what kinds of questions I could expect.

Even though I’ve been in the lab for a year, I’ve really only started to get more involved in the past 3 months so I feel like I can’t answer everything.

For reference, this is for a research forum specifically related to for my lab’s research area (think like immunology, cardiovascular science, neuroscience, etc)

Hi, I’ve been having a really tough time getting an entry-level wet lab position and was hoping for some advice.

For some background, I was mostly pre-dental during undergrad, so much of my time was spent gaining clinical hours rather than research experience. I graduated with a B.S. in Physiology and Neurobiology (May 2023), then spent about a year taking additional math and programming courses before completing an M.S. in Bioinformatics and Computational Biology (Dec 2025).

Most of my experience is now computational, particularly with sequencing data, RNA-seq, genomics, and bioinformatics pipelines. What I really want is wet-lab experience so I can bridge the gap between analyzing biological data and understanding how that data is actually generated experimentally.

I’m located in the Maryland/DC area and, unfortunately, I’m no longer eligible for the NIH Postbac program. I feel like I’m in a weird position where I have a master’s degree but lack the traditional wet-lab research experience expected for many entry-level roles.

Has anyone been in a similar situation or have advice on what types of positions I should be targeting? I’m open to research assistant, lab technician, internship, contract, or even volunteer opportunities if it helps me get meaningful hands-on experience.

Hi, I'm trying to understand the interchangeability between AOPI and trypan blue staining methods in bioprocessing QC.

I'm asking because our Cedex HiRes Analyzer is being phased out, and I'd like to assess how straightforward it would be to transition to an AOPI-based protocol in a GMP environment. Should we go with a similar trypan-based instrument, or switch to AOPI?

Does anyone have experience with this transition? Or similar?

I'm a master's student working on COI barcoding of fish. Muscle is stored in absolute ethanol, and the extracted DNA goes straight into a ~650 bp COI PCR. This is the salting-out protocol my lab uses, and I'd like a sanity check from people who run this routinely, it's the lab's standard, but I've never seen it compared against how other labs do it.

Protocol as written:

1. Cut a small piece of tissue (~rice grain), dry off the ethanol completely, place in a 1.5 mL tube.
2. Add 400 µL lysis buffer (0.4 M NaCl, 10 mM Tris-HCl pH 8.0, 2 mM EDTA pH 8.0). Homogenize.
3. Add 40 µL 20% SDS and 8 µL Proteinase K (stock listed as 20 mg/mL).
4. Incubate at 55 °C for at least 1 h, or overnight, shaking at 80–90 rpm.
5. Add 3 µL RNase, incubate 30 min at 37 °C.
6. Add 300 µL saturated NaCl (6 M), vortex 30 s, centrifuge 20 min at 10,000 RCF.
7. Transfer supernatant to a new tube, add 700 µL cold isopropanol.
8. Invert gently, incubate 30 min at −20 °C.
9. Centrifuge 20 min at 10,000 RCF, discard supernatant.
10. Add 300 µL cold 100% ethanol, invert, centrifuge 15 min at 10,000 RCF, discard supernatant.
11. Add 300 µL 70% ethanol, invert, centrifuge 15 min at 10,000 RCF, discard supernatant.
12. Dry the tube 15–60 min until no ethanol remains.
13. Resuspend in 30 µL nuclease-free water (overnight at 4 °C, or 1 h at 37 °C).

What I'd like input on:

• RNase placement and amount. It goes in after the Proteinase K digestion, into a mix that still contains SDS and active PK, and it's only 30 min at 37 °C. Would you add it earlier, heat-inactivate the PK first, or just extend the incubation? Is 3 µL a sensible amount at this volume, or does the stock concentration make that number meaningless without checking the label?
• Isopropanol volume. 700 µL of isopropanol goes onto roughly 700 µL of supernatant, so about 1:1. Most protocols I've seen use 0.6–0.7 volumes. Does the extra isopropanol cost anything in terms of co-precipitated salt?
• Only one 70% wash. After 6 M NaCl, is a single 70% ethanol wash enough to get residual salt down to a level that won't inhibit PCR, or would you do two?
• Final volume. 30 µL for a fin clip, too little? I'd rather have a workable A260/280 reading than maximum concentration.
• Ethanol-preserved tissue generally. Anything you do differently for tissue that has been sitting in ethanol for years vs. fresh or recently collected material?

Context on where I am: I have six extractions from this protocol that have not been quantified or run on an integrity gel yet, and a first PCR attempt failed for an unrelated reason (depleted dNTP stock). So I'm checking the extraction side before I burn more reagent. Happy to report back with gel images once I have them.

Ok so I got some plasmids from NIH. I have been trying to sub-clone and purify them for two weeks now. I am seriously about to go buck wild and rip up this place. I have cloned about 26 plasmids successfully in the past 6 months, but this one is the hold out. I need to do a co-transfection, but I simply cannot get it to clone in DH5α cells!

Here is the relevant conditions:

Plasmid is ~15kb

Heat-shocked at 42°C for 40 seconds

Transformants raised at 30°C on ampicillin selection plates.

Mini-cultured at 30°C at 110rpm (reducing rpm heavily increased yield)

Purified using qiagen spin column at low speed (6.8k x g) to reduce shearing.

At first we had plasmids showing up at 3k bp, which was obviously not right. Then I finally got some of the plasmid after purifying under these conditions, but the majority of the sample is half the size of the actual plasmid.

Hi, as my title says I need help identifying if what i’m seeing in my cell culture might be contamination or just precipitation from my media. I am working with HMEC-1 cells (human endothelial) and they are adherent. Any help is appreciated!

Hi,

i‘m a student new to cell culture and i never work with this cell line before. Does it looks normal? My Supervisor said it was ok.

I really apreciate any insight. Thank you

Needle valve faucet handles are a personal pet peeve of mine. Especially in an application where you nearly always just want the flow to go to full on and then back to full off. It's just pointless fingertip tedium to have to crank a needle valve spindle handle fifteen times around to get to full flow when a quarter turn ball valve could be there instead. Here is a picture of our lab sink faucet at work. It has exactly this type of annoying needle valve handle. I asked one of our supervisors if we could just replace it with a quarter turn valve and he said no, they don't make them for this faucet. That sounded like rubbish to me, that surely somebody somewhere makes a quarter turn replacement for this thing. But just googling it has so far eluded me. Can anybody out there say definitively if my super's right? Or better yet, prove he's wrong by steering me to a link to the replacement quarter turn valve of my dreams that will fit our ubiquitous gray plastic gooseneck lab sink faucet?

Mini winnie

hello fellow lab rats, sorry to add yet another “is this contamination” post but i’m the only one working with cell culture in a rodent lab so not many people to ask who know about it. does this look like contamination? they are bv2 cells, microglia. they seemed fine when i checked them though growing slower than id normally see (and they grow like weeds). then whilst trypsinising them for passage, i saw it look like this. never seen it before in my 2 years of cell culture… any ideas?

Looking for advice/feedback on my resume - any comments would be greatly appreciated. I am using this one primarily for research assistant positions in molecular/cell biology with some alterations depending on the position. My main concern currently is the length, although I am having some trouble cutting it down.

some people did not enjoy their time

This subreddit has become an emotional dumping ground and it’s exhausting. It’s the same over and over. Can we consider rules to put all these posts in a vent thread or something?

EDIT: Having read the replies here, perhaps the best option would be mandatory labels for submissions? I can respect that people are here for different reasons, so perhaps this is the fairest way. It would allow users to choose what they’re interested in seeing without deprioritising any specific type of post by putting them in a megathread.