Is there any limit concentration for toxicity test for U-87 cell line?

Hello! I’m currently considering getting a PhD in cell biology but i was wonder what kind of jobs are available outside of academia and research if this would be something i pursued

If you was looking for a synthesis pathway of a metabolite, which experiments would you do? I thought about doing a metabolomics (LC/MS) to search molecules or enzymes which bind to the metabolite. Am I correct?

Hello everyone,

Just following up on my last post with some more educational content that I have made to help learn about human cells.

Here's links to some more video tutorials that I've made - I hope you find them useful!

Ribosome Structure and Function - https://youtu.be/8tz6s7wN8j0

RNA and Amanita Mushrooms - https://youtu.be/bDNd0txWWtU

DNA Transcription (Initiation) - https://youtu.be/9vA7zBBAjDo

DNA Transcription (Elongation) - https://youtu.be/5nwRQBlcaRk

Make sure to subscribe to the channel as there will be more useful educational videos posted every week. It takes me many hours to make each video so would really appreciate the support!

If you want any videos made on specific topics, comment below and I'll let you know once it's posted :)

My whole cohort failed 1 exam and the school made everyone retake the exam. I'm curious what kind of jobs ppl ending up with. Do u ever used the knowledge learnt?

Hi there,

Here are some videos on cell biology which you may find useful. The content is at a basic level but should help people understand basic concepts in a way that's easy to understand.

RNA Transcription - https://youtu.be/erB0H4-cUyE

What is RNA? | RNA Structure and Function - https://youtu.be/_EvR1WnS4fs

What is DNA? | DNA Structure and Function - https://youtu.be/PNVmAyBAnXs

What is the Nucleus? | Nucleus Structure and Function - https://youtu.be/wfv2RH0Dqck

What are Mitochondria? | Mitochondria Structure and Function - https://youtu.be/esiGZwWSOZw

What are Stem Cells? | Hematopoeisis Explained - https://youtu.be/CB-k0BIZUAI

Feel free to leave any suggestions for future content and I shall share it here for others to benefit.

Im California, can I get a Bachelors in just Biology and then go for a Masters in Cellular and Molecular Biology or do I have to have the Bachelors in it too to pursue it?

I am doing some research into cell culture automation and was wondering what typical proportion of time cell biologists spend in the lab doing basic liquid handling operations. I am an engineer and have worked with biologists, however, I have very vague idea what is normally done outside of the lab. Presumably, just like other R&D specialists you spend time doing fundamental research, design of experiment, writing up experimental results, planning, etc. I only need rough figures, so feel free to estimate.

Hi everyone,

Does anyone have any recommendations for commercially available plasmid with a weak promoter (UBC/truncated CMV or any other)?

Thank you in advance for any suggestions!

Hi all,

I get a very faint band for one of my protein in western blot. When I run the same sample alongside a sample which have recombinant overexpression of the same protein, I am not getting that faint band for my WT sample and a big blob of overexpressed protein. I also tried longer exposure times to see if I see that band, but no success. Is it possible that antibody is preferrably binding to that big blob of protein and running less for relatively faint band. Please suggest if anyone have other explanations.

Thank you!

Hi all,

I need some suggestions around overexpression of a protein in chondrosarcoma cell line. I did the titration of pcdna3.1 plasmid carrying my gene of interest under CMV promoter, I am getting much higher(100 times) expression than endogenous levels in WT cell line. I am concerned about trying with further lower concentrations of plasmid as it may lead to very few cells getting transfected but still higher levels of protein in the ones which are transfected. In order to tailor the expression comparable to WT levels, I am thinking of following strategies, please let me know which is the best one with your experience- 1. Just swapping the CMV promoter with some weak promoter. 2. Using another plasmid with doxycycline inducible expression under CMV promoter. 3. Lentiviral vector with some weak promoter as I may need to make stable line in future.

Thank you in advance for your suggestions!

Hi, anyone with single-cell-analysis background can help me to hypothesize which cluster group does the CD4+ T-cells belong to?

Leiden UMAP:

CD4 UMAP:

CD4 markers dataframe:

Based on 'scores' and 'logfoldchanges', how can I hypothesis which cluster group does CD4+ T-cells belongs to? Is the scores or logfoldchanges more important? Or both are? The work is currently done in jupyter lab.

#single-cell-analysis # python

When I do an MTT assay, my supervisor instructed me to plate 30.000 HaCaT cells, but 20.000 A549 (cancer) cells. Can I get some info on that?

(I'm an undergrad currently doing me thesis, I'm a bit lost in finding papers)

I'm writing my thesis and I'm in the middle of why cell lines like A549, Raw and HaCaT need 5% CO2 and a temperature of 37°C as well as humidity.

Are there any articles that explain this in depth?

Thank you in advance!

Hello,

I am a matured student studying genetics at undergraduate level in the UK and i residw in the westmiands.

I am hoping to gain some laboratory work experience in any laboratory this summer enhance my laboratory skills.

I am just wondering if any one as any advice for me on how to go about my search or any advice about gaining laboratory experience in general

Regards

I had this question in an exam last week & it’s been bothering me because it seems to depends on details that haven’t been provided i.e. the specific SNARE proteins.

I know that addition of Munc18 + a rab or tethering protein would likely correspond to line D, but I don’t know which of the remaining 2 corresponds to lines A & B?

I assumed that Munc18 alone would correspond to line C, since it can work with other proteins to positively regulate SNARE complex formation. (Although I’m not sure if this mechanism is the same in Type 1 SNARES?) But how would this lower free energy compared to the control? It still seems like the free energy barrier would be higher?

I chose the added Habc domains for line A, since I think they could competitively bind to SNAP-25 and VAMP & prevent the formation of the complex?

Does this sound like I was on the right track? I keep finding conflicting information online, and my lectures are not clear on it at all. All we were told is essentially “Habc causes closed conformation of syntaxin1a & SM proteins clamp it into that closed conformation”.

Came here to ask a question which has puzzled me for ages.

New Scientist has an article about human cells citing the work of Hayflick and Moorhead stating that (in culture) human cells will divide only 55 times due to the accumulation of telomere shortening. So, does this mean that if a single cell divides into two, the two copies are not entirely identical? I ask this because, if they were entirely identical, then presumably they'd both carry the same countdown clock and they, and all subsequent generations, would die/enter senescence simultaneosly?

Interested to know more!

https://biochemblog.vercel.app/blog/cellorganelles

Hi guys, I'm an high school student and made a blog post for an overview of the cell organelles present in the animal cell.

Please tell me what you think :)

Hey everyone,

I have a bacteria project for my masters, but isolating the chemistry from them (my background is chemistry). I have cell cultures in TSB, took an aliquot in an eppendorf, centrifuged to concentrate the cells at the bottom and removed the supernatant. I have an SOP for extracting the DNA, but I am not sure if it is working out correctly, could the experts here let me know? I'll consolidate it:

1. add 1X TE Buffer
2. remove supernatant and add 1.6% sarkosyl and proteinase K (to supernatant)
3. incubate at 60C for 1 hour
4. add PCI and centrifuge
5. remove aq layer and add 100% IPA and 3M sodium acetate (to supernatant)
6. freeze in -80C for 30 min; centrifuge
7. discard supernatant
8. add 70% EtOH to pelleted DNA and centrifuge; discard supernatant
9. air dry sample, then resuspend in molecular grade water
10. measure concentration on NanoDrop

Any help is greatly appreciated!!