...look at structure, impediments, the representation of women on committees, and how the culture of the ... promoters and terminators, but then they aren’t sure how to convert their own coding sequence to the ...science based around those technologies in the near future. Science on Twitter Addgene: You are active on ...Twitter account recently inspired by an academic in literature who gave a talk to the women’s research network...

...proteins associated with the IRF-1 locus. Figure 1: Schematic of enCHIP. Image from Fujita and Fuji...affinity purification in situ of regulatory elements (CAPTURE) (Liu et al., 2017). This system utilizes dCas9...through proteomics and the chromosome conformation capture (3C) technique. In addition to 3xFLAG and biotin...FSHD), a disease for which there is currently no cure (Campbell et al., 2018). Find plasmids from the ...Central PMCID: PMC6286148. Liu, Xin, et al. "In situ capture of chromatin interactions by biotinylated dCas9...

...you don’t have plasmid ID numbers, or you aren’t sure if any plasmids from a paper have been deposited...plasmid search and found the results confusing, I’m sure you’ll find Addgene’s newly launched search engine...marker used to grow plasmid containing bacterial cultures Bacterial Resistance: Use this to choose the antibiotic...Editing brings us to a page with a basic overview and figure, followed by a convenient table of plasmids specifically...resources on the Addgene blog Learn more about other features on Addgene's website ...

...an antibiotic resistance gene is a mechanism to culture only bacteria which have retained a functional ...formation on agar plates, less stable at high temperatures and in acidic environments.   Carbenicillin   ...can be successfully used for plant and bacterial culture, but some types of bacteria, such as SHuffle cells...all-in-one antibiotic — it is usable in eukaryotic cell culture, yeast, bacteria, and even plants! The zeocin resistance...., & Brooks, G. F. (1986). Effect of storage temperature and pH on the stability of antimicrobial agents...

...applications. Read on to learn more about the current (and future) applications of lentiviral vectors! Lentiviral...popular for targeting and tracing cells in the brain (Figure 1). 2nd generation lentiviral packaging system ...to create the 2nd-generation lentiviral system (Figure 2). The HIV genes that do remain are very important... Dull et al. created the 3rd generation system (Figure 3), which differs from the 2nd generation in two...Gluckman E, Bushman F, Hacein-Bey-Abina S, Leboulch P. Nature. 2010 Sep 16;467(7313):318-22. PubMed PMID: 20844535...

...are required for replication and transcription (Figure 1). All viral genes are removed and replaced by...transcription, as it would for a full-length viral genome (Figure 2). While it is technically possible to encode ... luciferase. Firefly luciferase can readily be measured to quantify (indirectly) transcription performed...result in significantly reduced activity (4). To ensure that you end up with appropriate plasmid ratios... not essential for virus replication in tissue culture, and the human RSV leader region acts as a functional...

... grant readiness, identify grant opportunities, secure grant awards, and manage grant projects and funds...management, we track grant opportunity deadlines, and we ensure that grant reports are submitted and project evaluations...field of grants. It is no surprise that the U.S. Bureau of Labor and Statistics shows that employment of...program didn’t stay in one place within the city infrastructure. My career path changed when I was asked if...She and her team at J. Miller & Associates have secured more than $70 million dollars in grant funding....

...plasmids of these systems to Addgene in the near future. Addgene: Has your lab been working on any other...development in biology and hope to develop more in the future. Involvement in the Scientific Community Addgene...papers submitted to journals. In addition, it is my pleasure to interact with my colleagues in meetings of ...institutes in different places have very different cultures. In Basel, I felt that the time flows much slower.... I hope I can help change the situation in the future. Addgene: What advice do you give to post-docs ...

...can direct people to the AMA in the future and link to it in future blog posts on these topics. Others ... new questions and topics to write about in the future. The AMA was also cataloged for permanent reference.... I have a paid account which gives me all the features I need to save, organize and easily find the resources...on the internet or receive via email. The cool features are too many to describe here, but I have over...

...candidates have seen an actual chalk talk. Their first exposure to a chalk talk is usually their own. This is ...in the remaining time. The typical chalk talk structure is: What is the big question or mystery that ...livers for transplants. Growth of new livers in culture could resolve this shortage." What is the knowledge...Perfect your story and delivery. Design simple figures and diagrams to illustrate your vision. Practice...beyond the whiteboard. You may wish to create a sculpture or print a 3D model to better illustrate your ...

... neurological defects. Nature 550:398–401 . https://doi.org/10.1038/nature24058 Tillotson R, Bird A (2020...can find the reagents and resources they need to cure Rett Syndrome.  If you’re new to Rett Syndrome, ...walking. They may exhibit slow head growth, develop seizures, cardio-respiratory dysfunction, distinctive hand...gene therapies to clinical trials in 2021. Figure 1: Schematic of the human MECP2 locus and MECP2...

...both targeted alleles when gene editing.   Figure 1: Techniques to improve editing efficiency. ... RNAs wisely (and let them cut close).   Figure 2: Relationship between cut site to mutation distance... one or the other on each target site.   Figure 3: Optimal cut site to mutation distances for heterozygous... and heterozygous mutations using CRISPR/Cas9." Nature 533.7601 (2016): 125-125. PubMed PMID: 27120160...scarless genome editing in stem cells using CORRECT." Nature protocols 12.2 (2017): 329-354. PubMed PMID: 28102837...

...such as the brain, transparent so that the 3D architecture of a fluorescent neuron is visible. Chemical... al. "Improved tools for the Brainbow toolbox." Nature methods 10.6 (2013): 540. PubMed PMID: 23817127...neural circuit analysis in Drosophila melanogaster." Nature methods 8.3 (2011): 260. PubMed PMID: 21297619.... arises from expansion and dedifferentiation of mature pigmented melanocytes." Cell stem cell 21.5 (2017...of fluorescent proteins in the nervous system." Nature 450.7166 (2007): 56. PubMed PMID: 17972876. Pan...

...interactions found in nature. Once the bond is formed it is unaffected by changes in pH, temperature or organic ...Discovery of the biotin-avidin interaction   Figure 1: Biotin is enriched in egg yolk while its binding...affecting the avidin binding site. With all of these features it isn’t surprising that scientists began to exploit...with biotin ligase and monovalent streptavidin.” Nature Protocols 3.3 (2008): 534–545. PubMed PMID:18323822...

...formation requires a fully folded beta-barrel structure, followed by an intramolecular reaction that generates...shown) remains fluorescent in acidic organelles (Figure 1), showing that its acid tolerance is maintained...specific phosphoinositides (Varnai and Balla, 2008) (Figure 2). RNA production can be detected by using a MS2...specifically detects a specific RNA stem loop-structure (Querido and Chartrand, 2008). Finally, a nuclease-deficient...Quantitative assessment of fluorescent proteins." Nature methods 13.7 (2016): 557. PubMed PMID: 27240257...

...standardized leader peptide prevents secondary structures in the mRNA encoding your gene of interest from...in-vitro assays: Koch, Daniel, et al. "A pull-down procedure for the identification of unknown GEFs for small...monitoring sequence dynamics under various selective pressures: Miller, Corwin, et al. "Adaptation of Enterococcus...applications of PUF protein fusions reported in the literature include RNA detection, RNA cleavage, and translational...endonucleases with customized sequence specificities." Nature communications 3 (2012): 1147. Cooke, Amy, et...

...few hours at low cost with restriction mapping (Figure 1). Though some labs may prefer to send DNA directly...computer program) can then reproduce the DNA sequence (Figure 2). Accuracy and accessibility led to the widespread...reactions and may use Sanger sequencing. Looking to the future Southern blotting, restriction mapping and sequencing...sequence an entire genome for under $1,000 (10). The future of sequencing should bring efficient, low cost ...large amounts of DNA quickly and accurately. We're sure to see many improvements in a variety of biological...

...accumulated over the years using them. Indeed, the future of mouse work is bright as it is now easier than... 2013 (Cong et al., 2013)! Haoyi got his first exposure to CRISPR when he used this new technology to ...pyogenes. An alternative Cas9, from Streptococcus aureus, is as effective at editing mouse embryos as Cas9...appreciate the fact that it was first discovered in an obscure bacterium and that we still have much to learn ...Production of Gene-Modified Mice using Staphylococcus aureus Cas9. Sci Rep 6: . https://doi.org/10.1038/srep32565...

...to transfect, select, and isolate clones. This procedure is cumbersome and inefficient, as it relies on...promoter or exonic regions. If the cell’s chromatin structure undergoes significant rearrangement (such as when...specific genetic loci of integration [8].   Figure 1: Mechanism of the piggyBac™  transposase/transposon...cells with a Footprint-Free™ genome edit.   Figure 2: Editing DNA using the piggyBac™ transposon selection...26299960. PubMed Central PMCID: PMC4559351. 8. A. G. Uren, H. Mikkers, J. Kool, L. van der Weyden, A. H. Lund...

...mutagenesis. Chapter 6 - The future of FRET and fluorescent protein tools. Future developments in microscopy...the Addgene Blog Visit our Fluorescent Proteins featured topic page Read Joachim's Series on Choosing the...Protein Plasmids 101: Visualizing Subcellular Structures and Organelles Resources at Addgene.org Find...

...started with tissue culture The cultivation of mammalian cells in the lab, or tissue culture as it is commonly...antibodies or viruses. If you’re new to tissue culture, check out this video for some tips and best practices...practices for getting started with tissue cultures. To keep up on all of the latest protocol videos...

...recommend you read the article to ensure you understand the features of the plasmids, how they were created... to the PubMed Preferences and Filters pages, be sure to check out the other “LinkOut” badges you can ...Addgene Blog Get Tips on Keeping Up with the Literature How to Write a Scientific Review Article Tips...

...Massachusetts that means it’s time for near-freezing temperatures, our first of the season snow flurries, dusting...holiday season! It is that time of the year again: miniature #Christmas tree goes up at home!This year with...December 4, 2020 Here’s how to enter: Post your picture or video for either contest on Twitter or Instagram...follow us on Twitter and Instagram as we share pictures of our decked out spaces.       ...

... plasmids shipped daily may be easy to picture (see the picture of our shipping room above), but as Joanne...amongst the lab team, shipping team, and others to ensure that everything will ship out in time - from assembling...said during our celebratory lunch, it’s hard to picture one million plasmids. How much space would one ...

...lucky enough to have an imaging core or tissue culture core, you are looking at a process-oriented team...come across one. Also, remember, one important feature that all projects have in common - and by association...is a blog post, so here are just a few critical features based on my own experience: Leadership Few things...certainly necessary, but equally important is making sure all team members understand what roles they’re meant...well, you can learn from the experience and make sure your next team works better. In fact, don’t wait...

... of a scientist was like. The word “scientist” conjured up images of men like Albert Einstein and Bill... make decisions about what we want to do in the future. How do I get started with research? I had no idea...personal interests While it’s perfectly fine to be unsure, it helps to have a vague sense of what kind of..., even if a question is a very basic one, making sure that you have a good fundamental understanding of...goals helped me reflect on what I wanted for my own future. Informational interviews with people who have ...

...breakthroughs and undermine critical research infrastructure that has taken decades to establish.These funding...materials that might otherwise be lost amid lab closures or project terminations, ensuring that scientific...grant applications, addressing the new funding structures and indirect cost caps more efficiently. Offering...can navigate this difficult period together. The future of scientific progress depends on our collective...

...Exciting news! Addgene recently rolled out a new feature on our plasmid pages - links to articles citing...screenshot below to see what I mean. Additional features Once you've clicked on the "# References" link...article abstract and more. We've also included a feature to help save you from performing any unnecessary...domains" publication.  If you want to see this new feature in action, click here to explore the TetO-FUW-OKSM...

...Doudna and Eva Nogales presented the first detailed picture of Cas9's three-dimensional shape via x-ray crystallography...Cas9's structural details. "Understanding this structure may help us engineer around the current limitations...in Feng Zhang's lab, in a press release. "In the future, it could allow us to design versions of these ...University summed it up well in that New York Times feature when he said, “All of this has basically happened...

... useful tools because we can easily detect and measure the light they emit. Proteins that give off light...observed in the crystal jellyfish Aequorea victoria (Figure 1) in molecular detail. On the underside of the...this new green glowing protein could be used to measure gene transcription, as well as to track protein...GFP’s function. His lab described the protein’s structure and also discovered many mutations that alter ...

...Live-cell measurements of kinase activity in single cells using translocation reporters. Nature protocols...whose cellular localization serves as a proxy measurement of kinase activity. The key advantage of KTRs...nucleus, while FRET-based reporters can be used to measure kinase activity in specific subcellular compartments...

... a dish that features that ingredient. We just had a cook-off last week, and the featured ingredient was...Harry Potter-style.The shared lunches create a culture of openness and trust that comes from caring about...ll dedicate a post to these competitions in the future, and hopefully include some of our favorite winning...

...heart health. These are amazing advances that will surely save thousands if not millions of lives, but many...resources and promoting research for rare disease cures. Through their BeHEARD Award, the Rare Genomics ...better choose which genes and pathways to target in future small molecule screens. He plans to work with a...voice. Check back with Addgene often to learn of future rare disease research funding opportunities. References...

...values with each element. We also set out to make sure the page looked clean, professional, and modern....our website, or in social media, we wanted to make sure our friend matched our core branding. Blugene needs... our values of community and sharing.  One big feature upgrade? Blugene now has a full expression. You...Addgene's new search tool Read more about other features of Addgene's website Resources on Addgene.org...

...That includes keeping fees as low as possible to ensure that our materials are affordable but enough to...community-oriented values. Addgenies like the nonprofit company culture as well. We work every day to do a better job of... our core values. These accurately reflect our culture and outlook: Addgene increases productivity by...increase sharing in the community Addgene fosters a culture of respect, collaboration, and fun Addgene believes...

... first need to decide the best way to track or measure the process you’re interested in. Depending on ...examine, i.e. for an enzymatic reaction, you could measure consumption of a substrate OR production of a product...molecule biosensors Addgene has many sensors that measure the concentration of small molecules like arabinose...withstand a particular solute concentration or temperature? For further advice, see our posts on how to ...

... of the brain where DA quantitation is desired (Figure 1). The probe perfuses liquid, which causes the... the method is powerful and offers immediate measurements in a live animal that can be correlated to behavior...developed fluorescent-protein based DA sensors (Figure 2). The dLight1 dopamine sensor In 2018, Lin Tian...powerful new developments in the detection and measurement of neurotransmitters in various species and while...

...boxes). We get these boxes a lot when receiving temperature-controlled reagents. Expanded polystyrene will...company and it has to be packaging (no coated tissue culture plates, for example). Check the link for more details...or Axygen®, set up a bin or box in your tissue culture room for the wrappers and ship them back. Start...make a huge impact directly on waste, but also on culture. Encourage your workplace to become a leader in...

...Rodríguez et al., 2011).  Genetics of SPARC Figure 1: SPARC and SPARC2 use the GAL4-UAS system along...sparse (S) labeling of the cell population. Figure 2: Dense (c), intermediate (d), and sparse (e) ...gene expression and clonal analysis in Drosophila. Nature Methods 9:47–55 . https://doi.org/10.1038/nmeth...projection-specific brain-wide reconstruction of single neurons. Nature Methods 15:1033–1036 . https://doi.org/10.1038/...

...Ashley Waldron Organoids are three-dimensional cultures that mimic aspects of organ morphology and physiology... an organoid model (Sun, et al. 2021).  Figure 1: Example schematic from the Organoid EasyTag ...targeted cells that can be used to generate organoid cultures with a modification of interest. While the authors...other Cas12a nucleases identified, functions at temperatures up to 43 °C and can be used for genome editing...

... visualization. However, due to the nature of the size, structure, and other qualities of each tag class...individually isolate a protein, there must be a special feature about that protein that no other protein in the...for an extra clean up step of the tag – a handy feature. If you decide not to cleave the GST tag after/...

... lunch. There was something so special about her pure interest and curiosity to understand my work and...making of this photo!  Here, I was also given a mixture of support and independence that allowed me to ...on content strategy. I really believe that the culture at Addgene cultivates trust between coworkers and... more confident in myself, my abilities, and my future - something I didn’t expect from a co-op experience...

...for helpful information on the known features and relevant literature, or to decide which line is best for...cell line collections websites and catalogues Literature searches utilizing Google Scholar alerts and ...as commonly misidentified lines. Browsing this feature to know if your lines may be especially vulnerable...

...wanted to be. But prior to graduate school, I wasn’t sure a master’s was a good decision. So how do you know...means to gain employment, so it’s important to make sure a masters helps you get a job you want. But how ...with master's have Job descriptions often fail to capture all of the responsibilities of a role, so it helps...questions in this blog post can help you start to figure out if a master’s is a good fit with your interests...

...very painful FOP flare ups. Eventually, without a cure, AJ will begin losing mobility and not be able to...play like the other kids his age. A potential FOP cure But, there is HOPE. FOP is a genetic disease typically...solutions to treat rare diseases like FOP until a cure is available. Addgene and AJ's Avengers This year...knowledge of FOP, it’s biological causes, and potential cures. We encourage any researchers working on ACVR1 or...

...young scholars across Europe get introduced to the cultures and opportunities available to them at university...to students. Freshers fairs are also where many future scientists are introduced to the biology and chemistry...talks by prominent scientists, including Nobel Laureates, who speak about their work and current, trending...positions both inside and outside academia. Inspiring future scientists At Addgene, we’re incredibly glad to...

...Updated April 21, 2020 Picture this: you’re a busy scientist anxiously awaiting the arrival of the newest...signs each agreement just once, covering nearly all future requests/deposits. This reduces the processing ...between a TTO and Addgene, whose terms apply to all future deposits of materials with Addgene Master Reception...between a TTO and Addgene, whose terms apply to all future materials requests from Addgene MTA: material transfer...

...synthesis. They published their results in a paper in Nature Communications announcing a new bioinformatics ...good predictors because they are just that: rare. “Nature has many genes where there are not rare codons ...source genes from farther away in the tree of life." Nature communications 9.1 (2018): 4425. PubMed PMID: 30356044...to predict the lab-of-origin of engineered DNA." Nature communications 9.1 (2018): 3135. PubMed PMID: 30087331...

...readthrough mitigation. Nature. (2016) 534, 719-723. DOI: 10.1038/nature18308   Additional resources on...are unreliable for a variety of reasons (Baker, Nature). Tagging your protein allows you access to a ...Reproducibility crisis: Blame it on the antibodies. Nature (2015) 521, 274-276 DOI: 10.1038/521274a Arribere...

... both targets in your specimen. You scour the literature and find an ideal antibody against Protein A ...distinguish between your two targets.    Figure 1: In the isotype conversion process the variable...successfully visualize both proteins.     Figure 2: Two primary antibodies with the same isotype... the anti-Protein B antibody (Fig. 3)    Figure 3: Recommended secondary antibodies for rat IgG1...