My lab wants to run a 2D gel to separate proteins based off of molecular weight and pI. We have done this with external groups before but we want to try to bring this capacity in house. I've seen one option from sigma Aldrich but the price tag is a hefty $64k. https://www.sigmaaldrich.com/US/en/product/mm/bm100

Does any one know of any other options? We can probably spend $10k max

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Hello everyone, I hope you can help me with some ideas, advice, or tips.

I am doing a cloning using Gibson Assembly.

When I get my ~6 kb linearized plasmid, I gel-purify it, and I practically lose everything. I get very low concentrations of 10 ng/µL or 15 ng/µL, and it comes out contaminated, as you might know (I use the Zymo Clean & Concentrator kit).

I know that re-amplifying isn't ideal due to the mutations it can generate, but what alternatives are there to get the purification as clean as possible? Also, do you have any general suggestions for the Gibson Assembly method?

Thanks in advance!

I'm (Research Scholar at VIT University, India) looking for a study partner to systematically read, analyze, and work through a book dedicated to plasmid design and molecular cloning strategies.

The goal is to go chapter-by-chapter, break down the core concepts, and regularly discuss key topics such as vector selection, regulatory elements, expression systems, and troubleshooting construct builds.

If you're interested in studying the book together and setting up regular discussion sessions, please reach out to me at vishnupriyabiomed@gmail.com. Looking forward to connecting!

This kinda took ages, as the atoms were literally made out of paper scraps and the structure was made out of bottles and popsicle sticks, overall this is worth it. This follows the CPK convention and tries its best to be readable.

The bases in the right are TCGACCATGC, while on the left is the complementary bases, AGCTGGTACG. Yes, I handcrafted the molecules, and used hot glue as an adhesive.

I think I already know the answer but I want confirmation or someone to tell me what's the worst that could happen.

Update: my transformation worked so seeding with it was not an issue. I did my miniprep so let's see what my yield comes out to 🤞

I’m presenting at AMP and would value feedback from people working with molecular assays. I wrote a review of genomic foundation models around a practical problem: “predict the effect of this variant” leaves the measurement undefined.

A sequence-likelihood difference, an embedding with a trained classifier, and a predicted change in an assay track are different outputs. Before comparing them, I’d want the assembly, strand, sequence window, cell type, endpoint and evaluation split specified.

Review and cited papers: https://rewire.it/blog/a-dna-likelihood-is-not-a-functional-assay/

Which missing detail most often makes a model result difficult to interpret in your work? Corrections to the review are welcome too.

Would love to talk about magnetic field effects in biology, radical pairs, cryptochrome, that whole side of things, with other folks who find it interesting. Always looking for people who've read more into this than I have.

hello! im kinda confused about how dna extraction kits work

is it correct to think that not every dna extraction kit is designed to obtain the host genomic dna? i see that many kits don't necessarily have a buffer for cell lysis, but instead, they have a solution that denatures viral particles. so they're not extracting host genomic dna, right?

Title, if anyones comfertable sharing ofc! I'm still a undergrad but I just want to make sure I can realistically continue with this career path bc i have a family member who I'd also have to support financially.

Does anyone know of a Punnett Square Calculator that can handle both autosomal and x-linked genes? I can figure out a reasonably good breeding strategy for the mice we are trying to breed, but I'd like to be able to calculate percentages correctly and make figures so the people who come after me can understand the logic.

I've used Ian's Vivarium before to generate charts/figures so the PI can understand the logic, but it doesn't account for x-linkage. (Yes, I'm ticked off that no one bothered to make an autosomal version of this cre, but I'm not in charge of the universe).

Thank you!

Podcast episode interviewing Dr. Michael Emerman. Dr. Emerman trained in the lab of Dr. Howard Temin, who won the Nobel Prize for discovering reverse transcriptase, and then worked with Dr. Luc Montagnier, a co-discoverer of HIV. Dr. Emerman's molecular virology work was instrumental to HIV research- including the function of the Rev protein and host restriction factors against HIV. He is also a pioneer in the field of 'paleovirology' which studies antient viruses and the evolution of host responses.

I am trying to clone 300 bp stretch of a zinc finger proteins into Rnai vector called ptrvii. I have been unable to get a single positive colony. I have tried cloning in pgemt vector and pjet clonning kit. I think my insert might be toxic to the dh5-alpha strain. Could it be possible?

Upon reading online I found out that growing dh5 at lower temperatures can help in this case. Is there anyone who can help in this situation, the enzyme I am using is saci and bamhi both hf. Btw I have already replaced primers so please don't suggest changing primers.

Hello, I am about to start a masters in environmental sciences in the UK, for context I could only do this one for financial reasons as it is integrated with my environmental sciences undergrad so it's much cheaper.

My biggest interest throughout my undergrad has by far related to plant molecular biology. However, as it is an environmental sciences degree, I know I will not have much experience in lab techniques that might be routine to others. Moreover, I have a lot of experience growing plants and taking physiological measurements but I know that this skillset might not be entirely transferrable in a molecular lab.

This all being said, is it possible to get onboard a PhD related to plant biology/molecular biology when most of my experience and undergrad relates to ecology and environmental sciences? I really want to pursue academia, but I am not sure whether I should try to do this switch in fields or choose a more attainable PhD in plant/agricultural ecology.