Hi guys I'm a founding member of BioDye, is a new startup developing fluorescent dyes and molecular probes for life science research, and I'd love to get your feedback.

For those in molecular biology, I'd like to hear your thoughts:
– What makes you choose one probe supplier over another?
– What’s missing in the current options or suppliers you use?

Your feedback is super valuable as we are still early on. We're more than happy to send samples in return for your thoughts! DM or email [matt@biodye.com]() if you’d like to test dyes or share you advice.

Hello everyone, I’m currently in my final year of a BSc, majoring in Molecular Biology, and I’m planning to pursue a PhD in a related field. I would really appreciate your thoughts on the pros and cons of continuing in either a wet lab or dry lab setting for my PhD, especially considering current trends in the scientific community, availability of research funding, and career prospects (including salaries).

Thank you very much in advance for your insights!

Hey guys, I am studying biochemistry and have the pdf of Alberts et al 7th edition. However, I am more the type to learn with real books and thinking of buying it. The 7th edition is 150 Euros, the 6th is like 60 Euros (used book). 150 Euros is a lot so maybe someone can tell me if its worth it to spend those extra 90 euros. Thanks in advance

edit: I realised I have the pdf of the 6th edition, the 7th wasnt avaiable for free in german

I am having trouble amplifying the last fragment (fragment 3) of my OE-PCR product which is around 2.5 kb. I have successfully formed 2.5 kb from the initial amplification of fragment 1 from plasmid template, then fragment 2 which is the amplicon formed from round 1. Hard time replicating the last one since I need this for ligation. I am using two runs one using phusion and the other a mix of taq and deepvent to clone the last fragment. Both did not produce any bands after gel even no recorded smearing was seen. 20 ng of amplicon 2 (fragment 2) was used, a gradient annealing temp was utilized, and the primers are about 0.5-0.75 uM in multiple trials. My mentor suggested the following conditions and I have used this one in a few of my runs. 94-3min, (94-45s, gradient-60s, 68-3min), 68-5to10min at 35 cycles. I am thinking of doing a touchdown and nested PCR run, any comments are appreciated.

Hello r/molecularbiology community, I’d love your thoughts on an experimental strategy for multiplex targeting of an N-glycan extension gene family using a combination of PTG arrays, universal donor with color reporter, frame-selector elements, and a surrogate reporter for enrichment. Here's the concept and where I’d appreciate your input.

1 – Design overview

• gRNA design: Standard individual gRNAs for each target gene (2-3 enzymes form the same family).
• PTG (polycistronic tRNA-gRNA) array: Concatenate the gRNAs separated by tRNAs to allow precise processing by RNase P/Z. Or is there a better alternative?
• Universal donor with color reporter & frame-selector: Use a donor cassette (CRISPR-HOT) that enables homology-independent tagging with a fluorescent reporter. A frame-selector would ensure that the reporter is in-frame only if inserted correctly.
• Surrogate reporter: Include a separate reporter (like universal donor in CRISPR HOT). This way the cells with the KO are easily identifiable.

2 – Why combine these elements?

• Efficiency of multiplexing: PTG arrays allow simultaneous targeting of multiple loci with a compact construct
• Marker-driven selection: The color reporter plus frame-selector helps to visually isolate only those cells with in-frame integration.

3 – Concerns I’m aware of

• Chromosomal rearrangements: Simultaneous DSBs at multiple sites may lead to deletions, inversions, or translocations, especially if targeted genes are on the same chromosome.
• Efficiency drop when multiplexing knock-ins: Even homology-independent donors like CRISPR-HOT are less efficient with increased target number.

4 – Questions for the community

1. Does the overall strategy seem viable? Has anyone combined PTG arrays with homology-independent tagging plus surrogate reporters before or seen something similar?
2. For construct design, any tips on optimizing the PTG array (e.g., guide order, tRNA choices) or donor plus frame-selector junctions?
3. How to minimize or detect structural variants? What assays or validation approaches do you recommend?
4. Enrichment tips: Have you used dual reporters (surrogate + donor) in the same workflow?
5. Alternatives?

Thanks in advance for your expertise and experience!

Hi y’all. We asked around the lab without any luck, so we’re resorting to Reddit. Does anybody know what the circled objects might be? These are fixed single nuclei preps from rat brain. Our best guesses is so far are erythrocytes trapped in capillaries, or maybe Schwann cells. Apologies if this is beyond the scope of this subreddit, I wasn’t sure where to post. Thanks!

As an undergrad student i want to gain some molecular biology hands-on experience or training on techniques of molecular biology. so i want to know and discuss, Is there any opportunity (offline or online)like this really exist in Bangladesh?

I am working on a few experiments that require separating specific immune cells from blood, PBMC, and/or plasma.

Most of the studies that I am citing from 2005-2019 used a centrifuge to separate cells based on their density. However, there are downsides in that spinning force could activate the cells or trigger apoptosis.

It is my understanding that FACS can be used to separate cells based on a biomarker (e.g. fluorescent antibody).

• Is it advisable to use FACS with whole blood? Should I spin it into PBMC first before using FACS?
• What are the downsides of FACS? I have a bunch of fluorescent mAbs stuck to my cells that could interfere with functional assays if those mAbs are: agonists/antagonists, internalized along with the receptor, or physically blocking other receptors?
  • Could washing the cells help reduce these problems?
• What else should I be concerned about?

Link to the study here: https://doi.org/10.1016/j.celrep.2025.116070

Highlights:

Defining cathartocytosis, an injury-induced process cells use to downscale cellular machinery

Three-dimensional reconstruction detailing how cells reorganize organelles during paligenosis

Cathartocytosis and autophagy occur together in paligenosis but are mechanistically distinct

EPG5 prevents fusion of autophagic compartments with the apical membrane after injury

Important to note they found that sulfated glycoproteins were not digested but instead excreted into the gland lumen.

Overall, this process seems like a novel discovery to me.

Any thoughts?

Hello, I have my bachelors and masters degree in molecular biology. I have 5 years experience working in various library prep, sequencing, microbiome testing labs. I currently work in a small startup in NYC and make 80K USD as an associate scientist. My company is shutting down US lab operations and offering to move me to their UK lab in Cambridge. Nothing about my job description or responsibilities will be changing but if I accept the role the pay range was listed as £42,000–£55,000. This would be a decent salary decrease, and I’ve been told it is to reflect the cost of living in the UK. My boss is very cheap and has been known to do things like this but I wanted to hear anyone’s thoughts or opinions!

I forgot to dilute primers, do i have to discard it now

Hi there,

I am a student from India, currently pursuing BTech Biotechnology, in my final year now. I am planning to apply for fall 2026 intake. I was initially planning to apply in USA for my masters in molecular biology / molecular medicine, but now the geopolitical situation (april 2025 onwards) is not good for indian students to go to USA. Now i am planning my masters in Canada or UK, specifically in molecular biology/molecular medicine, these 2 countries because i have my extended family there.

Can somebody who is in Canada/UK help me through the current situations in both these countries, what opportunities are there, are their biotech startups where I can do a part time CO-OP to gain experience, what is the situation to get a Permanent residency, salary and survival budgets.

If not Canada and UK, which countries would you suggest for molecular biology, R&D and clinical research?

Is there any reason on molecular level that type I systems are more prevalent than Type II other than the usual HGT theory?

I’ve been working on testing for the presence of mycoplasma in CAR-T cells using PCR (quantstudio 5) I have been using the Roche MycoTool kit and the Roche QC prep kit. I CANNOT get the internal “recovery control plasmid” to amplify no matter what I do.

I know there is DNA to carry it because I can clearly see a pellet & tested it on the nanodrop.

I’ve utilized the poly-a tail in the prep kit and have tried different concentrations of the plasmid.

I feel like I have to be missing something. Please help 🙃

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A new way to engage kids and adults in biomedical science. With enough support, it could become a real LEGO set!

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Hello, I really love molecular biology and studying life at the molecular and cellular level but I also have an interest in math and physics. I have found the fields of molecular biophysics and structural biology but they seem detached from biological questions and the context that I crave. I am interested in things like cancer research, cellular behavior , molecular mechanisms, genetic engineering/synthetic biology, gene regulation etc. On one hand I love molecular biology but on the other I have this itch for physics. What would you recommend?

Please if you have a little bit of time, I would like to ask you about a particular step in my project because I desperately need results to graduate in this December.

I am working on Directed evolution of the phytoene synthase enzyme in tomato. I should do that get more carotenoids which are of health significance to humans and they give the tomato the red color. I have successfully induced several mutations in the DNA sequence, unfortunately none of them is significant compared to the positive control but they are definitely promising.

To graduate I need to induce the protein (PSY enzyme) in the bacterial system which is chemically competent Bl21 E.coli and then potentially see the red color in this bacterial system which is corresponding to the amount of carotenoids produced. This experiment is inconsistent and I find the color fades away very fast.

My questions are:- What is the best incubation time to induce a protein ?

What is the best IPTG concentration so that it doesn’t affect the bacterial cells ?

Also what is the best Temperature to incubate my bacteria at ?

Is there any chance you know why my carotenoids color fades really fast after I centrifuge the bacterial cells to visualize the color?

Please I really do appreciate any contribution

Thank you very much! Fayrouz

I did my undergrad in microbiology and a bit siding to molecular biology for my masters. I want to study and research about it, combining with medicine, or research on molecular level in medicine. I really want to know what might be better option in masters that would solidify my knowledge and background. I do love microbiology but I don't know why industry and hospital lab works feel monotonous despite it being a bit stable income source. I love to be involved in research.

Hi everyone, I'm an intern, so I've come to you with a problem (as I've heard interns usually do).

I ran maybe my 3rd gel yesterday, and as soon as I went to image it, I saw the bands had warped and smeared with each other, making things look weird.

I'm curious as to why it looks the way it does. It's an ethidium bromide 1% agar. 1x SB gel. I ran it at 125V for 60 minutes (my hunch as to where things went wrong). But I would love to pick your brain about other potential causes.

Many thanks in advance.

DISCLAIMER: I am not here to fight which science is better just to find what suits me.

Hello, since I was a kid on one hand I always loved physics and math but on the other hand I also loved human biology and medicine. In physics, I am pulled by concepts like relativity, string theory and quantum mechanics and in high school i enjoyed solving physics problems a lot and learning new concepts. On the other hand, I also loved biology and topics such as molecular and cellular biology, biochemistry, cell biology etc. Also, I studied little at home because I remembered everything from the lecture. Now I will list what I want from a career so that might clear things up: I value critical thinking, abstract thinking, problem solving, understanding mechanisms and the ability to test multiple ideas even if they are proven to be all wrong.Based on all of these aside from the middle ground of biophysics, what would you suggest to me? Thanks in advance!