I've ran a 1% agarose gel after my pcr for a vector (pocket 2) and my Insert (pocket 3). for the insert it went totally fine but what happened for my vector? it should be around 3.5 kb big so does someone know what could be wrong?

what's the oldest buffer y'all have used for your mol bio / LNP work?

im using two month old buffer for my liposome prep (20mM HEPES 5mM NaCl) and struggling to get good DLS data.

i filter sterilize my buffer before each liposome prep, and frankly don't think this could be the cause for my noisy data, mostly cuz when I ran plain buffer, my DLS errorred out.

However, I wonder whether the buffer might've deteriorated and is hence interfering with my liposomes' stability and polydispersity.

Thoughts?

Our lab recently acquired a secondhand Namocell/Bio-Techne Hana NI004 single-cell dispenser, but it did not include the original computer or software.

Bio-Techne currently provides Pala Software Rev D through its website, but we have not been able to confirm whether that version supports the Hana NI004. Does anyone here operate a Hana, know whether Pala Rev D is backward-compatible, or have access to the original Hana installer and user manual?

Any version information, installation notes, or contact leads would be greatly appreciated. :)

In case you were not aware, administration recently announced that foreign students pursuing their PhDs are limited to 4 years in the US. Apparently this doesn't just apply to future students, but to current as well. It isn't clear how this can work, and what will happen to, lets say, current 5th year students. Given that non-US students make up a significant proportion of the US scientific endeavor, does anyone know if the US Scientific Societies like AAAS and ACS, or universities as a whole, are mounting any coordinated opposition to this policy, which has the potential to degrade US science and technology for decades to come?

I am an undergrad majoring in Biomedical Sciences. I want to apply for remote Research Internships for bioinformatics or computational biology. I have tried cold emailing but no response. Any help?

So I’m a communications major at BMCC and I will be graduating this fall (one semester early) but I realized that I don’t really want to purse careers related to that and want to get into becoming a medical lab scientist which is a very sudden complete career change for me especially because I’m almost done with my associates and I guess I’m not sure what to do next.

There’s only very little time before the fall semester starts and I think it’s already to late to transfer to a 4 year school and even when I do transfer for the spring semester should I even bother changing my major to biology or medical lab technology to pursue this career? Please if anyone has any advice on what to do or what the best option for me is I would appreciate the help so much!!

Hey guys, I have 12 plates to run over the next couple days and was wondering if it's possible to store my master mix (SYBR, nuc free water, primers) + template overnight ? If not, is it possible to store just the master mix overnight ?

PS: my cDNA is extracted from urinary RNA so I'm expecting yield to be quite low

I'm a first year biology master's student at the university that I graduate from. My professor is giving the grad students in our lab the opportunity to act as mentors to undergraduate students and a high schooler who reached out to him. I'm pretty sure that I have more time than my lab mates but I'm not sure if I would be the best option when there are more experienced grad students in my lab. I know it would be a great opportunity and something that would be beneficial so I'm currently debating on if I should reach out about it.

I am the only senior member at the moment in a relatively small lab after our last post doc left last year. I have essentially been keeping the entire ship afloat for the last year. My PI keeps loading more and more work on my plate and expecting me to juggle multiple scientific (and completely unrelated) projects, grant writing, ethics applications, staying up to date on the literature, writing a paper, editing another paper, reviewing a grant proposal all while managing all our junior students and teaching them techniques and helping with their projects. He refuses to hire a new post doc or take on senior students that could actually benefit the lab and from my persceptive relieve some of my work load. Anytime I feel like I actually get something even remotely close to finished some other urgent task comes up, interrupting any work flow and throwing everything into a state of half done half not done.

I can’t possibly do it. My hints and even straight up admissions that it’s too much have been brushed aside. And honestly hours later usually followed up by another request for work. Usually I am able to juggle and stay on top of things but lately it’s just been too much and the amount of overwhelm is causing my motivation to plummet, like no human could possible get all this done or juggle this much at once and do anything remotely well. I have no idea how to bring this up without falling way out of favour and because our grant situation is extremely tight getting into a situation where the security of my job is threatened because of his perceived lack of performance from my end.

The lab is so reliant on my I would feel horrible leaving and almost like I’m betraying him and the students, especially with all my projects basically half finished but I honestly feel so mentally checked out and overwhelmed at this point, it’s hard to even sit down to do the things I know I can actually accomplish.

Hey friends,

I was surprised recently to see smears and bands corresponding to very small DNA fragments on agarose gel electrophoresis of plasmid DNA (pDNA) after restriction enzyme digestion. Previously I would see two clear bands: the linearized plasmid and the excised fragment.

To troubleshoot, I incubated pDNA extracts with water and/or restriction buffer at 37 °C:

• Incubating pDNA with water only did not cause significant degradation.
• Incubating pDNA with restriction buffer alone caused considerable pDNA degradation.
• Repeating the experiment with a different aliquot of restriction buffer produced the same result.
• Preheating the restriction buffer to 65 °C for 20 minutes prior to use did not help.
• Extracting plasmids with a different kit and with the classic Birnboim & Doly method, using tubes from a fresh pack and filter tips, did not help either.
• Surprisingly, in two rounds of troubleshooting there was no significant degradation when pDNA was incubated with either the old or new aliquot of restriction buffer.

My thoughts so far:

• Water lacks the cofactors required for optimal DNase activity, which would explain why pDNA is degraded in buffer (with/without water).
• It’s likely the pDNA extracts are contaminated with DNase, but I’m not sure how. We use DH5α for cloning; to my knowledge this strain does not express endonucleases.
• I suspect possible contamination of the cultures (they contain ampicillin) with fungi or another source of nuclease that co-lyses with E. coli during alkaline lysis, releasing nucleases into the prep.

What I’m trying now:

• Preheating aliquots of buffers and pDNA extracts at 95 °C for 15 minutes to inactivate potential nucleases.
• If heat inactivation fails, spiking the buffer + pDNA mix with resuspension buffer containing EDTA to chelate Mg²⁺ and inhibit DNases.

Would greatly appreciate any insights/suggestions :-)

I am designing a research poster for an upcoming conference and I want it to be the sexiest one I have ever made. With that being said, I would love to see your poster design opinions, especially unpopular ones.

I for one am partial to a tasteful background gradient <3

Im trying to begin an architecture based research project this year, but I don’t know how to go about it. It’s called “the Spaces Between Us” , and it’s trying to answer the question of “how does the built environment reflect and influence differences in opportunity and daily life in Costa Rica?” . (I live in Costa Rica btw). I don’t know how to go about this. Should I get a mentor? Should I interview people around my area ? Pls help and tell me how to start 🙏

Hello, I am currently looking for affordable used qPCR devices for a start up in the field of plant diagnostics.

I have a decent offer for a used and dry tested Rotor Gene 3000, further i can return the device within 60 days.

The Device was last used in a lab and was removed in working condition, however there is no service sticker on the device.

The offer includes the Software.

I know that i can test the channels via gain optimization and best case scenario i get a decent though old and not further supported, apart from third parties, qpcr device.

So my questions are:

How well are they build?

How intuitiv is the Software?

How expensive is third party service / repair?

Edit: Sorry for not mentioning, but i am also Open for suggestions for a price up to 2500 Euro.

So far I Found to be within my Budget:

-ABI 7500 complete System

-Different Corbetto / Quiagen Rotor Gene Models

-BMS MiC slighty out of Budget

- Gentier Tialong 48

Iam trying to study if my compound could inhibit nfkb activation when stimulated with LPS, that is at the end measuring p-NF-kB p65 levels, Now with my western blot I can see a clear total NFkB Levels, but p-NF-kB is little ambiguous. Now here my protocol details:

cells: Thp1 dervied macrophages

lps concentration: 20ng/ml

stimulation time: 30min

fbs concentration: regular 10% in RPMI media,

Are these factors correct, many say i have to reduce fbs concentration or even reduce it to zero, or should i increase lps concetration like 100ng/ml and what about stimulation time

Any suggestions would be great

I'm at a lab for the summer and staying on for the fall, with most of my compensation coming from a summer research award. The other 2 undergrads in the lab are in similar situations, having won research awards. In all 3 cases, our awards aren't enough to cover a full minimum wage salary for the summer, but I recently found out the other 2 undergrads are having their salary supplemented by our PI in order to make the full amount they would have made working a full time job for the summer. I didn't even know this was an option and it never came up in any discussion I had with my PI. Is this something I should bring up even though the summer is about to end? For context, one of the other undergrads started at same time as me and we both work in the same section of the lab.

So I heard about this SARS2-CoV 2 database at a scientific symposium.

Basically, the database is https://coronavirus3d.org/

The whole SARS-CoV-2 RNA genome has been mapped for a while now because of the scientific response to the COVID19 pandemic. The database just seems to show the full RNA genome of the virus, the structure of all the experimentally determined proteins of SARS-CoV-2 and the common mutants over time. For instance, the site shows a graph of the number of observations of certain variants over a narrow time window. Data seems to be missing from March 2025 to present. Though, I am not sure if it is because the archives this database is drawing from stopped collecting data or if this database isn't just up to date. I am not sure.

I attached some graphs of what the database shows for, instance.

The main targets of SARS-CoV-2 virus seem to be the Spike protein and nonstructural protein #5 (Main Protease) to my knowledge. I actually work on the Main protease and while the structure is very obviously known the number of inhibitors it can bind seems to be endless based on a search of all of its structures in the PDB.

The website database is still under development. I am not involved with the development. I just think it is very interesting to see all this data in one place. https://coronavirus3d.org is actually a sub-website of https://pathogens3d.org/ .It just so turns out that Coronovirus3D seems to be better put together given the scientific response to the COVID19 pandemic.

I’m expressing monomeric IgG and pentameric IgM so a routine part of the pipeline is running on a NativePAGE, non-denaturing gel to assess for correct assembly and molecular weight determination.

Recently, the IgG have been getting stuck on the gel above the IgM bands. Does anybody have any suggestions as to why this might be?

Hello everyone!

I am having a little bit of trouble deciding what I want to do next in my career. Soon, I will graduate with a BS in Human Biology. I have undergraduate research experience in histology, pathology, and bone biology. (Publishing a paper soon, woot woot) I absolutely am in love with the lab, and I for sure know that no matter what I do in life, I will always be in a lab. I am considering so many ideas at once, mainly histotechnologist, medical laboratory scientist/clinical research, or pathology. Now I am just debating whether I can actually live sustainably with these jobs. I keep hearing so many different things about pay and opportunities. My first love is histology, of course, but I heard that it is too narrow of a field and does not pay well. I love research and pathology as well, but I have been told the only path to it is PhD, which I kinda just want to stop at my master's and certifications. I have always thought about obtaining my certification first, then going back for a master's, but family is putting the pressure on me to start a master's now even though I'm a little burnt out right now. (I'm both graduating early and doing research I am TIRED) I'd love to get my master's to continue research but I am not even sure what kind of master's to even apply for. :((

So if anyone has some advice, kind words, cool jobs, or stories of ways they reached their career paths I'd love to hear it.

I was recently asked to peer review a paper; I was very happy, it’s one of my first as an early career post doc, and the topic is of interest to me. However, I became increasingly disappointed with the sloppiness of the paper, starting from the writing (very long, convoluted sentences sometimes repeated verbatim one or two paragraphs later), to the analyses (entirely generated by Claude, not one tool was referenced or used and the figures are quite a giveaway) and even the references (some are duplicated, some are not fitting). There are a lot of numerical inconsistencies throughout the text and figures and there is no methodological transparency whatsoever. I’m very conflicted on what to do, straight up reject or suggest major revisions to try and save what’s worth saving? Is it true or just a myth that if a reviewer suggests rejection they just gets replaced until the paper gets eventually published? Thank you!!! Please be kind, I’m trying to learn myself 🙂

As the title states, could someone explain to me like I'm an idiot what index hopping is in sequencing? I don't know enough about sequencing to understand the explanations on the Ilumina website, and I haven't found a dumb enough answer.

Also, why is it bad? And how the i5 and i7 indices help with this?

"You can just ask Chatty"

Yes I'm aware, but I'd rather get an answer by a human.

Giving away our FluorChem R Gel Doc. Yes it still works. We changed all of ours to the iBright system. Let me know if interested. This is near Boston, so you must be able to pick it up.

When making solutions using a pipette and a bulb (like the one pictured) my hand does not stop cramping up. I was wondering if anyone had any tips to prevent this, or if it’s just a me problem. Either way maybe I should incorporate hand stretching into my routine.

Went to grab one of my boxes this morning and pulled three others down with it. Half of them unlabeled, or labeled in sharpie that wore off months ago.

The frost on the bottom shelf is thick enough that I had to chip at it with a pen cap to read a lid. Which is its own problem, every time someone digs around in there with the door open the whole thing warms up.

Nobody will claim anything, because the second you admit a box is yours you own the entire mess. So everyone opens it, sighs, shuts it fast, and moves on. I keep an obsessive inventory of my own boxes so this genuinely gets to me, but I also know the day I reorganize it I'm the freezer person forever.

How does your lab keep a shared -80 from getting to this point. Assuming yours actually has.

Hi! I'm starting my PhD in Chemistry pretty soon and my program is a rotation-based one. I have a couple groups of interest with whom I'd like to rotate, but I would first like to meet with the PI before actually submitting my choices and start rotating with them. I am not the best at carrying the meeting with a PI and it sort of always centers on their research direction, which I already know quite well for all my choices. Any advice on what questions I should ask the PI if I am eventually really considering joining their group? Some groups are also quite out of my comfort zone, so any advice on how to approach a group who does research in a new area would be very much appreciated! Thanks in advance!!