Hi all!

Would anyone happen to have any strong feelings about a good C-Fos primary and secondary antibody in the rat?

I plan to do some IHC (perfusion fixed frozen sections) in the rat spinal cord and it seems like there are a lot of mixed feelings regarding the antibodies I’ve looked into thus far.

Any help is much appreciated!

So I know that artificial molecular machines are a thing, albeit limited in application

How do you like

make one

How do you put the atoms together

So I’ve been searching for a method to isolate EVs which are targeted to pancreatic ductal cells, is there a known way to isolate ONLY the ones that are targeted to there? Was thinking about binding antibodies and following up with ultracentrifugation but is there a better method?

Hello all,

For allele specific PCR I designed the forward primers for amplification of normal allele and for mutant allele and reverse allele but what I did was I designed the primers in such a manner that the forward primer began like from the beginning of the reverse strand and the reverse primer began somewhere in the middle of the sequence and not at the end of the sequence. My question is will this cause a problem in amplification? since the sequence amplified was also small

Hey everyone! Hope you're all doing well. I have 3 questions about primitive cells.

I was recently talking with someone religious about evolution and the origin of life. He used 3 arguments to defend creationism:

The necessary time to form a single protein (by random chance) is 1 in 10¹⁶⁴, which exceeds the Earth's age.

Nucleic acids, proteins, and polysaccharides break down in water, refuting the primordial soup theory.

Science can't prove if the first cells had RNA or DNA as their structural base. These compounds are also highly sensitive and wouldn't withstand the conditions of the primordial soup.

I don't have much knowledge on the subject, so any explanation would be welcome. Thanks :)

Im studying molecular biology and genetics. Its a 5 year degree here and im on my 2nd. The mandatory internship is normally on the summer between 4&5th. Our curriculum mostly consists of human physiology however plant and animals are also studied. My interest however is in immunology for the time being, but still early to make such claims.

Im thinking of doing a voluntary internship however the field is pretty limited in my country except for universities. I searched a lot of biomedical and biotech companies yet theres no openings for internships. I am thinking of emailing some professors in other universities about their researches that im interested in. Any advices on how to handle such things? Also any other internship opportunities other than universities and private companies?

hellooo i would like to ask what primer/s are usually used to barcode nematodes from fish? tyia!

Hi i have a new question. Right now our main goal is to optimize PCR conditions for target host genes in our project. If that is the goal, is it okay to just extract DNA and not RNA? We dont need to check at this stage what genes are being expressed or not. We just need to optimize the PCR conditions we are setting for the samples we will run in the future. Does it make sense to extract host DNA for this part? TIA

Im reading this paper below in which they show an increase in protein expression using certain regulatory elements. Id love to try this out, especially the tripartite leader sequence of human adenovirus mRNA. Ive combed throughthe referenced articles as well as searched for this sequence but i cant seem to confirm the correct sequence. Can anyone help point me in the right direction?

Thanks!

Mariati, Steven C.L. Ho, Miranda G.S. Yap, Yuansheng Yang, Evaluating post-transcriptional regulatory elements for enhancing transient gene expression levels in CHO K1 and HEK293 cells, Protein Expression and Purification, Volume 69, Issue 1, 2010, Pages 9-15, ISSN 1046-5928, https://doi.org/10.1016/j.pep.2009.08.010. (https://www.sciencedirect.com/science/article/pii/S1046592809002083)

According to this paper, a whole chromosome can be added or replaced by packaging single chromosomes into those "microcell" vesicles and fusing them with the recipient cells(that's what I understand). However, does the microcell only fuse with the recipient cell's membrane, or does it fuse with the recipient cell's nucleus as well? I have read over the paper but couldn't find the method of how the donor chromosome is delivered into the recipient nucleus. They only mentioned that PEG is there to help the microcell fuse with the recipient stem cells, and the Sendai virus can also facilitate cell membrane fusion.

If the microcells are fused with the nucleus, does the nuclei fusion only occur during cell division/mitosis? Can they do this with non-dividing cells like neurons? Is anyone here familiar with microcell able to answer my question?

Thought this would interest the upstream process developers of r/molecularbiology . Recipharm’s scientist will be presenting his protocols that increase their AAV titer and percent full capsids 2-10x across multiple serotypes today.

If you’re looking for a way to make your transfection more efficient, check out the webinar. Higher titers, higher percent full capsids, more stable complex formulation, lower cost per bioreactor run, etc.

Someone in our team did some western blots to visualize biotinylated proteins. So there was no 2nd antibody, instead STREP-HRP was used of course, sort of the same procedure, except it is not antibody. Anyways, he did a standard SDS page western blot, and got bands as expected. Now it comes, there are bands in samples where we do not expect them, but even more weird is that there are bands on the whole membrane (all around 55 kda, straight line over the membrane), even where no sample was loaded. And I do not mean a bit of spill over: even slots away of the last loaded sample. This happened a few week earlier to, same bands, same heights. They even are visible in the marker lanes. There is just no common factor between the samples and the ‘ghost band slots’, except the gel and the running buffers. We already changed buffers. Anyone has any clue?

Hello everyone,

I'm working on a python script that translates DNA and RNA strands into proteins, but I've stumbled across a hurdle which I can't seem to overcome. If you check all the reading frames, there might be multiple ORF's. Is there a correct way to always choose the right one? Internet says that the longest one is probably the best one. note: probably. Is there a set of rules to always choose the correct one?

Thanks in advance

I have a lab coming up where I’ll be given an unknown plasmid and will have to identify it by designing my own restriction digest procedure. I’ve been given a list of restriction enzymes I am allowed to used, and have looked on NEB for their respective protocols.

Since I don’t really know what to expect out of the concentration of my given unknown plasmid, I have no idea what concentration I should be calculating for when designing the restriction digest protocol. Any advice?

Does an open reading frame have to be a multiple of 3? Also, does it include the stop codon or exclude it? For example, say you have the sequence:

ATG GGC GGC AAG CCT CGG CGG AAG AAC CCC CAG GAA GGC CTG TCC TAA GTG AAA GGG

Would you say the ORF is (includes stop)

ATG GGC GGC AAG CCT CGG CGG AAG AAC CCC CAG GAA GGC CTG TCC TAA

or (excludes stop)

ATG GGC GGC AAG CCT CGG CGG AAG AAC CCC CAG GAA GGC CTG TCC.

Also, hypothetically, if the sequence was not a multiple of 3, and looked like:

ATG GGC GGC AAG CCT CGG CGG AAG AAC CCC CAG GAA GGC CTG TCT AA

Would the entire sequence be considered an ORF, since it starts with ATG and has a TAA end? Thank you.

Hello im currently 3 months away from starting college. Im very interested in genetics and other closely related science fields. Im wondering how hard is a molecular biology degree compared to a biology degree. I know that molecular biology is consistently rated a top 5 hardest degree but it also has lots i would like to learn about. I dont have much prior knowledge in biology outside of the core ideas. Would I be able to complete a molecular biology degree or would i be better off doing biology

Let's say someone had a disorder that is characterized by mitochondrial dysfunction. Normally, the body would respond to this mitochondrial dysfunction by attempting to boost glycolysis to make up for the energy deficiency. Part of this process would be using the PFKFB3 gene to boost F26BP levels, activating PFK, the primary rate limiting step in glycolysis. But what if the PFKFB3 gene was highly dysfunctional? What kind of workarounds would there be to activate PFK without the help of PFKFB3? Presumably something like pioglitazone or semaglutide wouldn't work because their actions actually depend on the functionality of the PFKFB3 gene. The psych drug Paroxetine supposedly activates PFK, but there's a line in the paper about it that suggests that its activation of PFK may depend on the "conformational changes" caused by allosteric activators of the enzyme. This is probably a stupid question but would it not be possible for someone to just administer the F2,6BP and call it a day?

I'm an undergrad writing a review article on cancer treatments targeting the DNA methyltransferase (DNMT1) enzyme, and I'm unsure about what to call the drugs when referring to them all together.

Some of the drugs I talk about are "DNMT-depleting agents" because they deplete DNMT levels through degradation or preventing protein transcription --> less DNMT in the cell.

Other drugs bind the enzyme and render it inactive, making those more accurately "DNMT-inhibitors". Some of these also have combined effects, both binding to inhibit AND marking for degradation.

While I haven't labeled any of the drugs as such, a related category would be "antagonist", but I see on some forums that "antagonist" is used to refer to molecules that block ligand binding to a receptor.

I use both these terms in my paper to refer to the appropriate drugs generally, but I worry that it draws attention to the distinction in mechanism which is not actually important for the purpose of my paper; what's really important is the consequence of inhibition/depletion/antagonism (being loss of DNMT action). I am tempted to just refer to all of them as DNMT1-inhibitors for brevity, but I don't want to be inaccurate. My question is: Is it important not to refer to a compound as an "inhibitor" if it doesn't chemically inhibit? Is there another word I can use that encompasses all of these categories?

Many people know about graphene, the wonder material that's 200 times stronger than steel. But what about Carbyne LAC (Linear acetylenic carbon) and Graphyne, which are significantly stronger.

You can find more information regarding these materials on the YouTube channel: Subject Zero Science, but the main takeaway is that Carbyne LAC is somehow twice as strong as graphene, and Graphyne is many times stronger than both materials.

My question is:

could these materials be theoretically infused with bone/skin/muscle/organs, would it be practical, and if it's not possible now, is it possible in the forseeable future?

Also, how strong would someone be if they were composed of 95% graphyne? Would their skin be so strong it would deflect military grade artillary and things such as missiles? Can organs be infused with graphyne to protect them? Would graphyne infused muscles give enough durability and strength to allow us to do feats like punch a tree down or lift a car?

I have a degree in Animal Science that I acquired overseas. I focused heavily on molecular biology, genetics and genomics. Although it was an animal focused degree I am fascinated by genetics regardless of the species. Now I’m in the United States looking to further my education and I don’t know where to continue. I’ve considered going into human genomic research. Is it possible to transfer over to human genetics? What is possible considering the degree I have and my idea for a possible shift in focus?

Hi everyone,

I have been using Superscript VILO for my TaqMan qPCR assays. I ran out of this kit, but have the Superscript II. Has anyone done any performance comparisons of two kits/enzymes if it aftect the TaqMan qPCR reactions?

Thanks

My daughter is wondering whether she needs a med school to become a clinical molecular geneticist? I know nothing about this and we have no one to ask. Anyone here that can answer our question?

What do you guys think of this post?

In short the author says the recipe is:

10% Store bought bleach (2L per 20L)
1% NaOH (200g per 20L)
1% Sparkleen or similar powdered detergent (200g per 20L)

https://pipettejockey.com/2016/05/06/make-your-own-nucleasenucleic-acid-decontaminating-solution/

How many ul of my 10um sirna should i use per well or a 12 well plate to do a knockdown with lipo2000?

I dont know the ug or pmol. I only know it is 10um stock.