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...use. This will prevent spills, but, if any spills do occur, they should be wiped with 70% ethanol immediately...morphologies, being familiar with these morphologies and doing a quick check under the light microscope will help...

...use But exactly what kind of editing are people doing with all these CRISPR plasmids (and libraries)? ...scientists use CRISPR technology? We think so — and so do our requestors! CRISPR plasmids in the Addgene repository...

...they are present at a very low concentration. So how do we make sure to catch all of these potential mishaps...expected hits, we always get hits to “random” genes. Do these hits always mean the sample is contaminated...users titrate their AAV vectors to find the optimal dose that allows for sufficient transgene expression ...

... = 500,000 colonies.  Pro tip! Since you probably do not want to individually count 500,000 colonies, ...did not result in significant recombination. If you do see recombination, we recommend ensuring the cells...

...proteins to pathways - Ras as a model When mutations do arise and cancer ensues, traditional chemotherapies...and thousands of investigators studying them, how do we ensure that the research community has access ...

...Gal Haimovich of greenfluorescentblog. Be honest. Do you really know how fluorescent proteins glow?   ..., you will not be able to use far red-FPs. If you do not have a filter that will pass blue light to the...

...Gamma-retroviral vectors do not have defined “generations” of plasmids, as lentiviral vectors do. For more about...

...offer tools to help with these decisions.  So when do you want to codon optimize your gene of interest?...useful in expressing functional proteins in hosts that do not naturally express that gene. The protein you ...

...negative control to determine whether your PIPs actually do interact. Choosing the right negative control is ...scenarios that call for a simple, yes-no answer — i.e., do two proteins interact or not? — FRET is preferable...endogenous PIPs should decrease the BiFC signal in a dose-dependent manner. This is straightforward in easy-to-transfect...easy-to-transfect cell lines where plasmid dosage can be tightly controlled. This is also theoretically...

...but orthologs like LwaCas13a do not require a specific PFS. Cas13 enzymes do not contain the RuvC and HNH...

...time, when scientists who support open science, but do not share their own data - which seems to be the ...members or colleagues: How much unpublished AAV data do you have sitting in computers and notebooks right... gene transfer to the rat CNS: AAV PHP.EB vector dose–response and comparison to AAV PHP.B. Gene Therapy...

... beads are particularly advantageous because they do not require centrifugation between washes which can...controls include knockout cell lines or tissues that do not express the protein. With this control set, you...

...gradients, practice making the iodixanol layers before doing your first purification. This helped me get a feel...to learn more about different ways to titer AAV. Do you have any AAV production tips or tricks? Please...multiplicity of infections (MOIs), to determine the optimal dose of AAV for my cells of interest. Every batch of ...

...interference), and CRISRPa (activation) libraries that do just that (Doench et al., 2016, Sanson et al., 2018... identifying strong hits in genetic screens, they do have several limitations. CRISPRko results in a complete...

...life. That’s convenient for researchers who want to do experiments in complex eukaryotic cells, but would...to express your protein of interest. However, you do still need a species-compatible promoter for expressing...

...five of these Cas proteins, while type I-C Cascades do not contain Cas6. Regardless of the subtype, the ...to the multi-Cas mechanism. These benefits of Cas3 do come at a cost — the previously mentioned issues ...

...cancer via insertional mutagenesis (which HIV does not do). Furthermore, molecular biologists working with ...in order to understand what these vector particles do once they come in contact with target cells (Fig....

... team at Calico Life Sciences recently set out to do just that — by developing a new fluorescent protein... MagLOV raise broader questions like “How and why do fluorescent proteins respond to magnetic fields?”...

...components can interfere with conjugates. Make sure to do your research before conjugating at the bench.   ...to distinct, non-overlapping epitopes so that they do not interfere with each other. The detection antibody...

...fluoresce, the Jaffrey lab asked:  Could an RNA aptamer do the same thing? Using the HBI derivative 3,5-difluoro...recognition of ions or particularly small molecules that do not generate an immune response. Aptamers can be ...

...fluorescence microscopy (Dani, et al., 2010). How do you overcome this resolution limit? Electron Microscopy...Prakash K, et al. 2022). So what’s a researcher to do if they want to understand the ultrastructural relationships...

...expression. To design a viral vector system that could do both, Addgene depositor Didier Trono and collaborators... lentiviral system (Figure 2). The HIV genes that do remain are very important for viral production: Gag...

...Gene X in the untreated cells—but can we be certain? Do we know whether the plasmid delivered into the cells...negative controls that produce a similar product, but do not target an endogenous gene in your experimental...

...of necessary data requirements and likely will not do so (calling it a “sisyphean task” given the endless...Firstly, we rely on many other labs to teach us how to do experiments and understand their data. Likewise, ...

...other aspects of molecular biology. “What I'd like to do with the plasmids from Addgene is have the students...

...quantification so that you can focus on what you want to do with your plasmid without spending time and resources...

... Michael Früh submission, which we have entitled "Do You See What I See?" I was bored this weekend while...

...expect given the excitement surrounding anything to do with CRISPR, our most popular post in July (with ...

... 2025. Catalog items are updated more frequently. Do you have suggestions for other plasmids that should...

...Cancer. 2003 Jan;3(1):11-22. PubMed PMID: 12509763 . Do you have suggestions for other plasmids that should...