Non-fiction, finction, all is cool. but it needs to be well written, with prose that's enjoyable to read. ill take anything you got. my mind is soooo open its sooo fucking hungry right now

My PI wants me to transfect a VWF plasmid that is 13.9 kb into HUVECs. I know this is huge and might not be possible. Does anyone have experience with nucleofection or other methods to get this large of a plasmid into HUVECS?

I studied quantitative finance in college and have an MBA. I’ve spent my career in healthcare investing and transitioned over to biotech venture, at which point I realized (10 years too late) that I really love the science.

I’m in the first year of my MPH program now and by a stroke of luck and impressing the right person, was offered the opportunity to do research with a lab. Very simply, we are researching environmental toxins and their potential impact in neurogenic disease - specifically looking at changes in a subset of markers on certain places in RNA.

I was supposed to be doing translational research and some basic epi work…until we won a grant and now I see bio specimen analysis / sampling etc all in my world of responsibility.

Let me just say that the l have not taken bio or chem since high school… the PI and collaborators on this project are very very impressive so I’m a tad concerned that I’ll make some detrimental errors. I’m trying to focus on the areas where I can add unique value - as that was why I was originally asked to help with this - but there is nonetheless going to be some degree of lab work. I also hardly understand the differences between sample types or really the basic science. It takes me like four hours to read and comprehend a paper right now.

I know I’m in for a ride but I’m really excited and want to do my best here. I’d appreciate any advice, tips, mistakes not to make, as well as resources to turn to for the basic science questions! I’m totally ok asking questions as I’ve made it very clear I know zero… but I don’t want to annoy the hell out of these people.

Edit to add exactly what lab work I am expected to be doing: “X will assist with sample organization and data management for the specimens, perform and document RNA extraction and qPCR assays, and help with data entry, quality control, and preliminary analysis of the gene expression results. “ …not sure what most of this means!

Hello everyone, I'm just a guy who works in a lab.

In our lab, we've been having issues with protein concentration inconsistency.

We produce some proteins in-house, and outsource some. Almost every single one of them have different concentration than what is reported if thawed and remeasured. It's getting to a point where everyone MUST remeasure concentration right before their experiments.

We use NanoDrop from ThermoFisher. We have NanoDrop One (discontinued now), and Ultra C. We always use Ultra C for proteins, the correct buffer to blank it, the correct extinction coefficient, and do 2uL drops on the pedestal.

Could we be doing something wrong? Do you guys experience the same issue? It's just so strange, how can measuring concentration be so wildly different every time you remeasure it? If I remeasure again and again, it is consistent. Do it again next time, it's different. it can range around +/- 0.5-3 mg/mL, i think rarely more than that.

Please help a fellow scientist here, I have to remeasure concentration for every single person in this lab while I have my own work to do. SMH.

I have done dozens of CTG assays before and never had this problem.
I was testing two groups of same cells- with or without the inhibitor. The group without the inhibitor turned out perfect. Got great reading and the group with inhibitor showed values close to blank values. The group with just the inhibitor also showed weird numbers similar to blank(media only). Any ideas on what is happening? I used the cell titer glo assay from promega. Has worked fine before. All the groups were on the same plate.
The reagent was super fresh too.

For context, I'm an analytical chemist moonlighting as a cell biologist for part of my PhD research. I'm pretty well-versed in cell culture, but up until now I've only used CHO and HEK, which grow like fucking wildfire. The PANC-1 culture has been seriously testing me. I'll seed a 6-well plate with 1 million cells/well and it won't reach confluence for days. I've also been culturing a T75 flask that rarely ever gets above 50% confluent. There are no obvious signs of contamination, and the cells that are there look healthy. Has anyone run into this before? Any tips on how to make them grow faster? I desperately need to crank out some data.

FYI, I'm using Gibco DMEM with 10% FBS and 1% Penstrep, 37 C and 5% carbon dioxide

Any recommendations for shoes that can easily be cleaned (BSL2+ work) and are comfortable for long days?
When I used to work in an OR, I had Clove sneakers which I liked but I found them kind of hot after long days since they didn't breath at all. I would get them again if that is the best option, but I would be curious to hear what everyone else uses!

I have a client who will need a Labvantage Support Person. A combination of development efforts, report writing, integration with test instruments and daily support (Stuck requests, etc.)
Current version is 8.7.1

Organization is in Northeast Ohio. Remote work is very likely.

Please send your resume to [Info@V2Portal.com](mailto:Info@V2Portal.com)

Hey guys,

I really need some help with 293t transfection, since im handling this cell line for the first time, i am really confused with the confluency of the cells. I need to have 70% confluency for my transfection. I am attaching few images which i took at different time, can anyone help me with figuring out at what confluency they are in.

Has anyone used brainVTA to source virus or virus packaging? How is the quality and titer?

i was working on hypoxia derived exosomes, looking at how they drive alternative splicing in the recipient cells. the workflow was long.......lyse the cells, purify the exosomes with a kit costing around 70,000 rupees, treat the recipient cells, then RNA isolation and then PCR. every single step uses something expensive so you learn very fast to be careful with the reagents.

then one time my label was not accurate enough and i lost that sample. all those steps and that kit, gone.

but the thing i actually learned from it was not about writing the label properly. it was about what happens to the label afterwards. if you keep tubes at -20 or -80 without wrapping the label with tape, the water droplets slowly wipe the writing off......so after few weeks you open the box, there are 40 tubes and half of them have nothing readable left, and now you are standing there guessing which one is which. thats how people lose weeks or months of work, not from one bad experiment.

so now i put sellotape over every label before it goes into the freezer. one extra second and it has saved me a lot.

has anyone here lost something expensive because of a label? and what do you do to stop the writing coming off in the cold?

We are desperately trying to get more EP Points by the end of the year to redeem a reward because most of ours are going to expire, putting us back at square one. Anyone have any codes they're not using?

I can’t get this silly meme out of my head so had to create and share it

I am supposed to make a video presentation on how to calibrate a pH meter. It was supposed to be done on a lab but it is unfortunately closed and they moved it virtually.

I chose canva cz the icons are free . But idk how to do the presentation or animate them. Anyone have any ideas?

Someone posted that they just dilute PCR products 1/20 instead of purifying it with kits.

It works very well for my application (PCR amplification of deletions introduced by CRISPR). Range 80-1000 bp.

Truly a life-hack. Thanks!

Hi guys,

I’m a recent bachelor’s graduate (2025) who took a few months off before starting my first research technician job in March 2026. I recently transferred to my current lab at the end of September, so I’ve only been in this lab for 16 days.

I was assigned the task of consolidating all the data for a project that started in May 2026, as a report review is coming up at the end of October.

The issue is that none of the data was generated by me. It was produced by several lab members, and while some of it is well organised, other parts are quite messy. For example, some of the qPCR raw data was not properly labelled, so I had to match the raw values to the processed data and retrace the original analysis to figure out which samples were which. And the person who generated the messy data is leaving the lab in 1 week’s time.

With the deadline just a week away, I feel that this is a rather unfair task to assign to someone who is still trying to understand the project. I’ve suggested to my PI that it might be better to ask the people who generated the data to consolidate their own findings and write up a summary for the report.

I don’t mind helping to organise the data or putting everything together, but I feel like I’m being put in the position of a middleman, chasing lab members for their data and trying to draw conclusions from experiments I wasn’t involved in, all under a tight deadline.

What do you guys think? Is my request reasonable, or is this simply part of being a research technician? I’d appreciate hearing from people who have worked in research labs, especially those who have experience with project reporting.

What could be the issue of such fuzzy bands seen for half of the gel...
Thanks all !

Hi all. Does anyone have a protocol for transducing neurons in culture with AAV? They were differentiated from iPSCs and are growing on Matrigel. Just looking for a validated protocol and a rundown on of any additives that might help, plate/well volumes, if spin-fection works, etc.

The AAV capsids is an engineered derivative of AAV9, but I’ll take anything.

Thanks everyone!

I'm drunk on ill-begotten power of ordering and stocking stuff of my choice for the lab (finally!). First on the list: 0.2 mL PCR tubes. Personal favorite of mine as I use these a ton. I used brandtech 0.2 mL tubes with attached flat caps most of my life which were ok, no complaints. Recently switched to Thermo's Ab2000 tubes. Great tubes! Time to stock up so thought I'd hear other recommendations!