/r/labrats
Got lab stories? Experiments gone wrong? PIs gone AWOL? Need help with your experiments? Share your stories!
/r/labrats
Currently building a product in automotive manufacturing to pull relevant info from scattered data sources, curious if it'd be needed in biotech or if it's even a problem here
I now feel intimidated by Figs in this format, always wondering if they are ai generated
Hello everyone!
I am a new PhD student and will be working quite extensively with single cell RNA sequencing analysis. I have no experience in this so my PI would like for me to attend a workshop/course to gain some background in it. I will be using the data to map developmental trajectories and understand immune cell cross talk. Does anyone have recommendations for courses or workshops that I can take a look at?
I need to buy some biotin for use in proximity labeling experiments. It turns out the cheapest biotin from our supplier is a pharmaceutical secondary standard, intended for use as a reference standard when measuring biotin concentrations. But wouldn't it still work fine in research?
Just wondering if anyone here has done an APHL fellowship before? if so what sort of resume/CV format did y'all use for your application?
I am trying EMSA using native gel. This is stained my sybr safe. I ran ladder as well to monitor if my gel is running fine (1st lane). Why do I see kind of straight line on one side of the band. It is in ladder as well as in probe and probe plus protein lane. I am running gel at 15 mA. I don’t think it is fast. Need help.
From South Africa
About to use the last of my lab's mounting media (Vectamount Permanent Mounting Medium) but it seems to have been discontinued. 90% of our needs are just H&E, with occasional IHC, IF, and even more rarely trichrome. Any recommendation?
I have been running gels for over a year now targeting two genes. From top to bottom: Ladder, cytb gene, ND4 and then repeating cytb and ND4 for the next 4 lanes. I used 3 samples and successfully amplified ND4 for all of them, but obviously I didn’t get cytb at all. I ran a 1.5% gel at 105V for 60 minutes. ND4 is around 700bp while the cytb should be 200-300 bp. Why didn’t I get cytb at all and why is my ND4 so smeared? Also, why does the ladder also look like shit? It’s brand new.
These questions might be stupid, but I am becoming extremely frustrated and would love any advice I can get. Thank you in advance🥺
But I like to lurk this subreddit.
Oxygen tank empty in the procedure room? I'll change it right away, but can you guys meet me halfway by maybe learning how to use the regulator?
Thanks!
Should i still proceed if there are bubbles in the cuvette after i added the competent cell and plasmid. There was a spark and the reading is 1.8(i assume this is a good sign), then i proceed to incubation and plating. It turns out there was no colony.... So if there are bubbles in the cuvette after i added all solution, should i still proceed? or should i just discard the cuvette?
Hi everyone. I need to store an aliquot of compensation beads for 48 hours . Is it okay to keep it in a foil covered eppendorf at 4c or better to keep in a foil covered FACS tube with Staining buffer at 4 C? Would this detrimentally affect the beads ability to bind to the antibodies when I add I use them after 48 hrs ?
Thanks for the help .
I'm using the Qiagen soil DNA extraction kit. The final step, when I centrifuge to remove the DNA from the filter, requires the column to be put in an elution tube. I leave the caps of the elution tubes open because there doesn't seem to be any other option, so every time I centrifuge during this step half the elution tube caps rip off and I have to carefully pipette my DNA into another.
Is there a solution for this? I'm placing the caps so that they rest toward the center of the centrifuge. I haven't tried resting them the other way (caps up toward lid) since that seems like it would be worse.
Any help is greatly appreciated!
I am studying a SNP in an intron of an 11 exon gene. Someone who worked on the project before me and who has since left designed 2 minigenes to study the effect of the SNP on the splicing of the exons before and after it.
One minigene was the genomic sequence from ex8-ex11 and the second minigene had an ATG and then the same genomic sequence. We do not see the same affect with both minigenes. One shows alternative splicing and the other shows a reduced amount of RNA product.
I'm a bit confused on what to make of the data. I'm not sure why they even did the ATG model since it seems like a typically minigene should just be the genomic loci of interest. But there are also reporter minigenes that would need a start codon to produce the fluorescence reporter. I was told it was to drive it to translation where it undergoes NMD hence the reduced amount of RNA we see in RT-PCR, but i don't think that is a fair conclusion to make since it is just a region of the entire gene.
I'm struggling to reproduce the alternative splicing result and questioning which minigene actually produced it now. No one in my lab aside from me studies splicing, so no one has been able to help me understand why use both minigenes or if one is better to use over the other. Any thoughts?
I had 2 groups of 3t3l1 cells: GFP- control GFP-GPNMB Under the microscope , I’ve estimated that GPNMB showed like 40% transfection efficiency based on number of green fluorescent cells and GFP-control showed like 60%.
When I ran a western blot my results showed high intensity of GPNMB on the control and lower in the GPNMB-GFP cells . I’ve been very cautious with these samples . Am I going crazy?
I've been scrolling through this subreddit for a while and I've noticed a lot of people getting trinkets or even food items when ordering stuff for their lab. I'm curious about this so is this normal, how does it work, does every company do this or only a special few?
For a master's thesis, the total wordcount has been told: 13,500 words (excluding abstract, tables/figures, references).
They haven't really said how much should go into each section and when I asked they said it depends. So how much of the word count should be devoted to each section? Is 20% for the introduction too much?
Hi everyone how are these surgeries that induce illness in rats ordered by hardest to easiest to perform? Surgeries: cholestasis, septicemia, varicocele, menopause induction.
Idk it just gives me mixed signals
Before i went back to school, I was really excited about becoming a nail tech. (hear me out, they make about $100 an hour and are their own boss). i also grew up love doing my own nails and intricate designs.
but when reality kicked in, i decided it would be better to have a career i can fall back on if anything ever went wrong with being a nail tech.
so i went back to school. at first, my family (who only think i will ever be successful if i have a college degree) said they would help me out financially to pay for school since i won't get help. but after my first semester, no one wants to help me out anymore. i live on my own and have a part time job (that i am sick of as i've been there the past 5 years and other than the fact that we can do overtime, there are no benefits or room for growth but i do get paid $21 an hour and get $2 raises every year). my friend who is in the same school has been telling me for months that i would love the medical lab technician program. and after being rejected for the radiology program, all the other majors didn't interest me, except for the med lab tech program since it's behind the scenes.
the first semester, was hectic as our school is severely underfunded and understaffed so both classes felt like we were chickens with our head cut off. and if i'm being honest, the only reason i passed is because i was able to cheat on my midterms and finals. i did not learn anything.
i'm now in my second semester and the classes have gotten worse. my microbiology professor quit the first week and we had a substitute for the first month. they were able to find a professor that has no recent experience of micro. my other class is so disorganized and has had no syllabus since day one because they decided to redo the class when the semester started. so this is looking like another semester where i will not be learning anything.
yes, i can cheat my way through. but what happens when i have to take the ASCP exam??
then my friend finished the program and passed the ASCP said she was having a really hard time finding a job. she ended finding one by luck that pays well, but now im second guessing putting myself through a financial burden just to get a crappy paying job. i'm honestly sick of studying science and i don't see myself going for my bachelors. and i'm scared if i end up messing up one day in the lab and losing my job...
i feel so helpless. idk what to do. a part of me just wants to stop going to classes and just take a nail course and be a freelance artist.. but i'm so far in already.. i only have 4 classes left to graduate and if i drop now, i'll still have to pay my full tuition.
i've had really good luck in getting a research scholarship last year and a fellowship program this year.
I would just love someone's guidance on working in a lab as a lab tech, good or bad? I'm about to be 25 and i hate the thought of just wasting more time of my life, not doing anything, staying at a dead end job.
just wanted to add that this dilemma has been making me so suicidal and i just want to quit life period. i'm tired of everything.
There have been discussions here from time to time about work with animals, like why it is necessary, how the actual work is being done, ethical concerns both from the public and from inside the lab, and so on. However, most of the discussion and sometimes concern is centered around common model mammals or sometimes teleosts. What about non-model organisms? I am mostly asking about reptiles such as lizards, snakes, turtles or alligators, non-xenopus amphibians, and secondarily about arachnids, non-cephalopod mollusks and non-teleost fish? First of all, how common are those animals in research? They are hardly ever discussed here, so they should be very rare. Also, is it true that studies with non-model taxa get less easily accepted? How are those animals sourced? There is no standardization like in mice and rats. In many studies, I read that animals were sourced from the wild and in others from breeders. Do pet breeders easily give animals for research? How common is for the research facility to have its own colony?
Then, where are those animals being used? Also, is the work on them similar with other lab animals? Are they usually long-term or single use? How the lab personnel interact and work with them? Have you noticed any sign of intelligence or communication in some of those animals?
What is being done for painful procedures? How do lab personnel feel about operating or euthanizing those animals? Are people going to skip the rules, just because they are not endotherms? Is the attitude of the people different if the species is native, pet or invasive?
Has the public ever been concerned about those animals? I don’t mean the animal rights extremists, just average people who may ask about how rodents, dogs and primates are treated. Given that so many people give herps much less attention than even common pest mammals and small teleosts I doubt it, although I will still make the question.
Everybody here seems to be in biology/biochemistry/bio-something. From the weird western blots (don’t really know how western blots work other than proteins move), to mouse/gnat/bacteria not working how it’s supposed to, to using micropipette tips in the wrong order I have no idea what you all are talking about. I use pipette tips to pick my teeth because we have 300 cases of them that’ll never be opened.
BUT don’t get me wrong I’m here for the bitching, I ABSOLUTELY love the bitching. I have no idea what most of you do and would lose my mind trying to figure it out. Generally speaking many of you don’t know what I do. But we all can gather over shitty PIs, freebies from the mail (thorslabs sends “lab snacks” if you ever need optics), and ridiculous memes. I think that’s beautiful.
Also fuck biology and organic chemistry can burn in hell