... Watson and Javier García-Nafría from the MRC Laboratory of Molecular Biology. Plasmid cloning is an essential...with successful recombination reported in all laboratory E. coli strains tested so far. The use of this...low cost and can be immediately adopted by any laboratory. All procedures are performed using the same... Watson and Javier García-Nafría from the MRC Laboratory of Molecular Biology. Jake Watson is a postdoctoral...postdoctoral researcher in neurophysiology at the MRC Laboratory of Molecular Biology. He is currently investigating...García-Nafría is an investigator scientist at the MRC Laboratory of Molecular Biology. He is interested in neuronal...

...early employees spent this remaining time by elaborately pranking each other, dressing in costume, and...frequently ask the question, “What’s your favorite part about working at Addgene?” Most Addgenies reply with an... confident that their teammates and project collaborators have completed their work thoroughly. It’s kind...rose-colored-glasses-esque picture of how everyone is kind and collaborative and all we do is have fun all day long, whistling...megaphone, or your company’s intranet. Don’t forget about the power of simple word of mouth - be sure to bring...company.        Additional Resources Read other posts about life at Addgene!Read more of our careers posts! ...

...on becoming grant writers. When we asked others about how they became grant professionals, mid- to late-career...late-career grant professionals have similar stories about a time when their passion for an organization or... logic models, established partnerships and collaborations, and commitments to and proof of compliance...grants. It is no surprise that the U.S. Bureau of Labor and Statistics shows that employment of grant writers... skills & passion  Skills such as research, collaboration across multiple disciplines, writing, project...development strategy builds on the steps listed above and formalizes your development. You may consider...

...Learn more about AAVs Resources on the Addgene website Visit the AAV Data Hub Learn more about the viral...resources available from Addgene. There’s a lot to talk about! And by the way, there will be viral vector jokes...injection site, and incubation time. Learn more about the Data Hub in this blog post. Alongside our efforts...SfN2019, stop by booth #2163 to say hi and learn more about Addgene’s viral service. Additional resources on...

...disadvantages of this overall rewarding role. Learn about Other Addgenies and Their Careers in Our Career ...general experimental guidance, and resolve complaints about products. As you might expect, different companies...order to enhance the OD signals and then complain about high background. In many instances, variations ...protocol. And customers may tell all sorts of details about the application, just not the ones that matter. ...communication, and is particularly excited to hear about novel developments in the fields of gene and cellular...Academic v. Non-Academic Career Decisions Learn about Science Career Options Explore Tips for Technical...

...Learn All about Mammalian Vectors Read up on CRISPR Delivery Systems for Mammalian Cells Learn about the Dangers...Misidentified or cross-contaminated cells have been known about since 1966 when Stanley Gartler reported that most...there is a loss of reputation that can impede collaborations, especially with industry. Today we also have...The LGC Group also provides standards and has laboratories to carry our independent quality control for...culture contamination." Archives of pathology & laboratory medicine 133.9 (2009): 1463-1467. PubMed PMID...

...fraction (<1%) of authors have asked for advice about the scope of the policy. End of story? Not quite...the data policy was raised by researchers worried about the effort to make the underlying data easily available...making data available, but there remains concern about the extra work this may entail.  So if access to...be archived? Simplifying data access Some of the above mentioned issues could be mitigated by on-line services...him on Twitter @jwoodgett.     More Discussions About Data Sharing: PLOS Data Sharing Policy Discussion...Scholarly Kitchen) Read more Addgene blog posts about scientific sharing ...

...Promoter of the arabinose metabolic operon Inducible by arabinose and repressed catabolite repression in the ...Browse All Plasmids 101 Posts Learn about Mammalian Vectors Read about Reporter Gene like Luciferase and...and operators (response elements). Promoters are about 100 to 1000 base pairs long and found upstream of...this list is a great place to start, the tables above do not delve into the tissue or development-specific...

...screenshot above). Don’t see your favorite organism listed? Help us serve you! Let us know about other organisms...useful plasmid that’s not in the collection? Tell us about it with our Suggest a Plasmid Form! Finally, if... if you would like to get the word out about a particular microorganism, microbial technique, or microbial...microbial technology, sign up to write about it on the Addgene blog. ...

... Read about frankenbodies, an anti-HA scFv-based probe for fluorescent imaging. Learn more about antibodies...Köhler who developed hybridoma technology. But what about single chain fragment variables (scFvs)? scFvs weren...from inclusion bodies, it’s time-consuming and laborious.  To avoid this, one or more of these approaches...antibodies  Read about another reagent sharing platform for antibodies, the Developmental Studies Hybridoma...Hybridoma Bank Learn more about the SunTag system which uses an scFv to amplify the fluorescence intensity ...

...occurring regions in the 5’ UTR of mRNA that bind metabolites, leading to conformational changes that control...Spinach-based riboswitches can be used as stable metabolite sensors for imaging in live cells. By placing...placing Spinach downstream of a riboswitch, metabolite binding to the riboswitch instead activates Spinach ...Dimerization of Broccoli increases its fluorescence by about 1.8-fold, and may be a better choice for imaging...modification that reduces nonspecific binding with RNA by about 7x). Mango fluoresces nearly as brightly as Spinach...Additional resources on the Addgene blog Learn more about aptamer fluorophores Investigate other fluorescent...

...thiamine metabolism Requires exogenous thiamine for growth ara disruption of arabinose metabolism pathway...little prokaryotic minions that carried out the labor-intensive process of replicating your new plasmid... Phage resistance gal Mutation in galactose metabolism pathway Cells cannot grow on galactose only ...repressed more   LacZ β-galactosidase activity abolished   lacY Lactose permease inactivated, lactose...plasmid stability recBCD Exonuclease V activity abolished Increased plasmid stability, reduced recombination...

...Addgene Blog Learn about Plasmid Awards to Researchers Working on Rare Diseases Read About Some of Our Outreach...Allied Sciences in Ghana. Read on to learn more about doctors Bengyella and Duedu and how they plan to...He hopes to teach his molecular biology students about the functions of these powerful tools and to use...China Additional Resources on Addgene.org Learn About Addgene's Mission Check Out Our International Shipping...

...Addgene blog. In fact, we have so many blog posts about careers that we’ve compiled our top career blog ...wait until you need a job to meet people and learn about what's out there.”- Joanne Kamens, Executive Director...school, I came across Joseph Campbell, and read about his "follow your bliss' mantra and never forgot ...Usually when I see inspiring artists who are thinking about getting into the arts (or any career to be honest...follow your passion. Do what you love. Don't worry about the rest.”- Jason Snair, Product Designer On the...

...ways scientists have adapted them for use in the laboratory. The discovery of GFP The story of how fluorescent...particularly interesting. Although humans have known about bioluminescent organisms and their characteristic...sequence. Soon after that, in 1994, Prasher’s collaborator Martin Chalfie expressed GFP in exogenous organisms...studying it grew. In particular, studies in the laboratory of UCSD biochemist Roger Tsien resulted in much...for you, so if you’re interested in learning more about fluorescent proteins, I invite you to download our...

...promote scientific sharing. Listen to learn more about Darrell, his research, and his focus on open science...Medicine and in the Department of Pathology and Laboratory Medicine. He attends in the Medical Intensive...NIH-funded Principle Investigator in the Kotton Laboratory and is the founding Director of the Center for...Podcast.     Additional Resources from Addgene Learn about Retinal Gene Therapy from Connie Cepko Find Plasmids...Plasmids from the Kotton Lab Learn more about Stem Cells ...

... on CRISPR Mouse Models Learn about Homology Directed Repair Learn about Cas9 Ribonucleoprotein (RNP) ... a variety of species, including the standard laboratory mouse. It’s also made targeted insertions relatively...process of conventional gene targeting is lowered to about 2-3 months. But certain models remain more difficult... a mouse embryo to create chimeric mice. This laborious process takes ~8-10 months. With CRISPR (B), you...directly into a zygote and obtain a knockout mouse in about 3 months. Image courtesy of Wenning Qin. Single-stranded...

...USA-Europe professional collaborations, setting up bases and contacts with Spanish laboratories to start working...where individuals can exchange ideas, network, collaborate, and innovate internationally. Why an International...needs, and came from the universities mentioned above. The initial IMP cohort of 78 mentees included: ...    Other Resources Visit the IMP website Learn about the Mentor Circle Program at Brigham and Women's...Networking for Scientists Series Read our Blog Post about Choosing a Mentor      ...

...well-integrated into our lab lives. Read on to learn about some creative approaches to lab sustainability, ...for rerouting waste from the landfill, but what about all the manufacturing steps that happen before you...ACT label, which is an eco-nutrition label for laboratory products that emphasizes Accountability, Consistency...away your FACS machine after a single use. So how about washing and reusing pipette tips in a high-throughput...Additional resources on the Addgene blog Learn about recycling initiatives for the lab Subscribe to the...

...attracting so much attention. When I spoke with Daniel about the interplay of these two ideas he said “science... scientists sometimes forget how to communicate about their work with non-academic people.” Daniel went...perception, which is ironically what science is about. For me it is essential to stay in touch with the...unique samples, meaning that we know very little about these specimens - especially those which are now...Additional Resources on the Addgene Blog Learn About Experimenting in an Open Source Lab Resources on...

...a summary snapshot without telling you anything about outliers or data spread within those data sets. ...the rules” and which we have certain information about. The assumptions and requirements that need to be...Quantitative If your data doesn’t fit the requirements above, a non-parametric test may be the way to go, which...only performing the comparisons you really care about. Family-wise error is the probability of a false...rate than the multiple comparison tests described above. Violated assumptions and statistics Statistical...not meet the assumptions for the tests described above (normal distribution, equal variance, and independent...

...Addgene blog Learn about the INTEGRATE system for bacterial genome engineering  Read about the Sleeping Beauty...DNA transposons are most commonly used in the laboratory for genome manipulation. When transposons are...Applications of transposons Now that you’ve learned about some of the popular transposon systems, let’s take...2020) Jump around: transposons in and out of the laboratory. F1000Res 9:135. https://doi.org/10.12688/f1000research...Beauty DNA transposon Learn about the piggyBac DNA transposon ...

... of Bioinformatics reached out to us to tell us about a resource called Cellosaurus, a growing cell line...all the information in the resource.  This is a labor-intensive project; scripts can be used in some cases...preferences to help you identify proximal lines you care about. If you have cell lines without a certified origin... Or finding errors in the available information about a cell line you’ve used? Submit your manuscript ...distributor. A success all around! If you’re excited about this resource and want to learn more, check out ...

... for bag check was definitely worth it! Hearing about his passion for global health and the various initiatives...with fluorescent microbes. Check out her animation about the Hawaiian bobtail squid!  Continuing the art...talked to some scientists who stopped by the booth about amazing work already represented in the Addgene ...opportunity for Addgene to let microbiologists know about our mission of open science and resource sharing...community, contact [email protected] to find out more about our free deposit and distribution service! Did you...

...It's been about 14 years since I last attended a Keystone Meeting – far too long. Holding these meetings...CRISPR technology in just 2 years and excitement about how much more there is left to do with the characterization...downstream output. Tuesday morning's talks were also about circuitry and synthetic biology, and I found myself...plasmid sharing to those who were anxious to hear about what was coming next. I also got to show off our... updated Synbio page to people who were curious about these newest features of our website.  I want to...

...new plasmid tool from the past year? Let us know about it in the comments or shoot us an e-mail at [email protected] and maybe we can write a blog post about it! Split Cas9 system The Cas9 protein is composed... lentiviral expression (MuLE) system from the laboratory of Ian Frew simplifies the process of creating... seem preposterous. Luckily, the Pierre Neveu laboratory designed a new iterative, chaining-based cloning... to improve on the Arch indicator and their collaborators in Robert Campbell’s lab carried out five rounds...improved light-switchable bacterial TCSs Jeffrey Tabor's lab has developed new synthetic biology tools by...for controlling gene expression. For more details about these light-switchable TCSs, read our blog post:...

... also be collecting feedback from the community about how the antibodies work in various applications,...antibodies are plasmid-based and recombinant. Learn more about recombinant antibodies here. Finally, Addgene is...against PSD-95 from the Trimmer Lab. Information about the antibody, the target antigen, and performance...blog and Antibody Guide, which include information about immunofluorescence, conjugates, isotypes, and more...learn more or get started.  If you have suggestions about how to make the resource better or ideas for new...

...Twitter, you might have seen the video of her singing about the paper recently published by her lab and Mustafa...professor at the University of Freiburg. This paper is about a new tool called BLADE (blue light-inducible AraC....   Inspired by @UriAlonWeizmann, here my song about our paper recently published in @nchembio Title:... based on the concept of bacteriographs created about a decade ago. To create these images, the scientists...Additional resources on the Addgene blog Learn more about inducible promoters Visit the Plasmids topic overview...

...living as a scientist in Beijing, talking to people about taking beautiful photographs of their studies, reporting...to friends or dates or colleagues we tell tales about parts of our lives that tend to have a story structure...informational interview question: Interviewer: “Tell me about a time you had a conflict at work.” Interviewee:...situation down by opening a dialogue specifically about the project and contributions, rather than getting...defensive or getting into ‘he said; she said’. We spoke about it for a while and he eventually calmed down, apologised...

...focus on molecular pathways, I found myself curious about other routes – In terms of both types and topics...not resist applying for. Not that I knew anything about being an editor - that I learned on the job. The...science? Look at our graphics and design editors. How about condensing a week’s worth of the world’s scientific...the scientific publishing world and debate issues about how to improve the scientific publishing process...science communication professionals who are thinking about, for example, how to use new technologies to reach...

...resources on Addgene.org Learn about Addgene's viral service Learn about viral production Find viral protocols... By now you have probably heard about Addgene’s viral service but did you know that Addgene also performs... approach can be adapted by vector cores and laboratories alike as an easy and inexpensive method to improve...Effect of medium osmolarity on hybridoma growth, metabolism, and antibody production." Biotechnology and ...

... your science with others.    My favorite quote about science ever comes from an interview in which someone...storytelling business like to say, science is all about navigating conflict. In literary terms, conflict...find that once I sit down and talk to a scientist about their research, these conflicts come out quite naturally... a simple question, like “Have you ever thought about how X works?”, or even by introducing a person that...devices, we sometimes forget the most important thing about science: It’s cool. Like bowties! Using science ...

...our Summer SciComm Series, where we’ll be talking about science communication, best practices in the field...research.   This summer, we’re making time to talk about my favorite thing - science communication (scicomm...endless.    Activity Example audience  Talking about your career as a scientist underrepresented minorities...students in AP biology classes Creating a podcast about genetics using dog breeds as an example adults ...get into the more exciting parts - learning more about science communication itself! See you next time!...

...section-specific comments, I re-read what I wrote about the paper after my initial read, adjust my language...wish for The last thing that I wanted to mention about the peer-review, is that as a peer-reviewer, you...each peer-review, I try to mention what I liked about this paper and recognize the hard work that the .../visualization using R/Shiny. She is passionate about open science, BIG data analysis and sharing whatever...review article Resources on Addgene.org Learn about depositing plasmids pre-publication Find molecular...

...We are excited about our new partnership with Allele Biotechnology which allows researchers to deposit...blue-shifted compared to its parent protein CFP484. Collaborating with scientists from Florida State University...mWasabi plasmids here! One of the most notable things about these proteins is that they are monomeric fluorescent... fluorescent protein blog posts Find blog posts about choosing fluorescent proteins for your research ...the Addgene blog Resources on Addgene.org Learn about the Michael Davidson fluorescent protein collection...

...frequency of use in black. This paper talks more about the development of the De-jargonizer.   Academic...with others which makes it easier to work with collaborators. Zotero is a desktop based program that’s compatible...episode 2 "Organizing Literature" : I found out about Zotero here Dear Grad Student podcast episode 2 ...Programming, & Productivity": This podcast talks about Mendeley, Zotero, and other citation managers and...the pomodoro technique": This podcast talks more about using the Pomodoro method in grad school Additional...

...the Addgene blog Read more about COVID-19 databases and resources Learn about Addgene's work to share plasmids... availability of this strain from The Jackson Laboratory. Originally developed and characterized by Stanley...mouse strain is regaining popularity. The Jackson Laboratory, a nonprofit research institution, has been able...clinical manifestations, transmissibility, and laboratory studies of COVID-19: an observational study. ...

...distinguish between DNA and RNA. See our protocol about using UV absorbance to quantify DNA Fluorescence...absorbance based methods, this method doesn’t tell you about contaminating proteins or chaotropic salts. Capillary... instruments are available. Some last reminders about quantifying DNA In choosing your DNA quantification...Additional resources on the Addgene blog Read more about molecular biology protocols and tips Find the best...best way to elute and store your plasmid DNA Learn about the different grades of plasmid DNA preps Resources...

...resources on the Addgene blog Learn about company culture at Addgene Read about other lunch activities at Addgene...scientists bring biases to research and don't collaborate with others to bring diverse perspectives. Everyone...Science is a terrifying and equally good wake up call about the science of race."     - Joanne Kamens, Executive...easy reads, some are hard. Some I’ll never think about again, some I loved so much I’ll force them on everyone...

... and we thought it would be a good time to talk about the amplification and verification processes we ... DNA. These reactions should allow us to obtain about 100 to 1,000 bacterial colonies per individual plasmid...from the count_spacers.py file from Feng Zhang’s laboratory. There are a number of other analysis scripts...you have any tips, input, or stories of interest about library use, let us know in the comments! Additional...NGS Sequencing to Validate AAV Preps Learn More about Pooled CRISPR Libraries Resources on Addgene.org...

...Bachle and Ina Ersing Seeding Labs and Addgene collaborate to provide scientific resources, equipment, and...discovery, and synthetic biology. Read on to learn more about the five awardees and their promising research projects... to the Addgene blog for updates and future collaborations. Many thanks to all of the awardees, all who...Additional Resources on the Addgene Blog Learn More About the Addgene-Seeding Labs Plasmid Grant Check Out...Expression Systems Synthetic Biology Resources Learn about Global Plasmid Distribution ...

...during the day? We asked three lab heads how they go about selecting new hires. This is followed by some perhaps...visits the lab for a full day, gives a 1 hour talk about his or her interests, and spends one-on-one time...current members to share short-term goals and chat about awesome science. Important factors in the search..., the lab has a reputation for not entering collaborations. Alternatively, perhaps the lab publishes and...Medical School) for taking the time to speak with us about adding new scientists to their labs. Read the Other...

...We also want to tell you about things you didn’t know you wanted to know about, like research breakthroughs... want to know more about! You can also suggest blog topics or even reach out about writing a guest post...technical skills, and experiences from thousands of laboratories that makes up our Help Center, blog, website...

... Learn more about research applications for antibodies in Intro to Antibodies Read about the Developmental...antibodies in your experiments, you may be curious about all of the different kinds of antibodies that are...Disadvantages of monoclonal antibodies As mentioned above, genetic drift is one concern when using hybridomas...Hybridoma Bank, a repository for hybridomas  Learn about monoclonal antibodies for immunotherapy Resource...

...reason behind this isn’t really hard to guess. Think about it like winter tires in different conditions: the...stable folding and efficient functioning. But what about placing the mesophilic hosts at 0°C? That won’t ...them which help foster a more productive and collaborative environment for research.  We’ve also involved...understanding of the societal ramifications of laboratory science, and how it may best be leveraged for...re always delighted to engage with potential collaborators and interested audiences. Please visit our team...

...Cancer, Inflammation and Immunity and learn more about “... recent findings in basic immunology, cancer-immune...curing cancer in cross-disciplinary projects and collaborations. Summarized by Professor Michael Karin from...This was followed by a talk from Professor Karin about alcohol and obesity as driving factors for chronic...got a chance to talk with Dr. Friedmann-Morvinski about tools used to analyse the tumor microenvironment...community, contact [email protected] to find out more about our free deposit and distribution service! References...

...often relies on antibiotic resistance. But what about identifying clones that contain your desired insert... restriction digest. The key is to be strategic about your enzyme choices. You’ll want to pick enzymes... for your PCR.  Find the details and learn more about colony PCR in this blog post.   Figure 2: ...Sanger sequencing or the other techniques mentioned above. In molecular biology, cloning a gene into your ... and tips Resources on Addgene.org Learn more about Sanger sequencing Read the Molecular Biology Reference...

...in Huntsville explains it, a major goal in his laboratory is to understand the function of the non-coding...protein tagging. We spoke with Mendenhall and Myers about their new CRISPR tagging system, which they’ve now... antibody against them. There’s also a question about how specific those interactions are. This is solved...This design cuts out weeks of waiting and hands on labor. That’s a huge deal. With Addgene, now these can...system or who use and adapt our reagents to think about adding them to Addgene’s special collection of CRISPR...

...to understand the basis for cardiovascular and metabolic human diseases. They do that by studying patients...disease-associated genes. Addgene spoke to Musunuru about how the new kit works, the research the kit now ...number of genes involved in cardiovascular and metabolic diseases. We are knocking out genes that have ...to examine the effect of a gene on cholesterol metabolism. The TALEN kit is quite useful in generating ...disease modeling in stem cells. Addgene: Tell me about the history. How had investigators done this kind...

...information about the contents of the autophagosome nor does it provide information about what is being...higher pH of the vacuole. Read on to learn more about prior methods for studying autophagy and how Rosella... (as opposed to giving more fine-grained detail about what proteins or organelles are being degraded) ...autophagy, but, like the protein degradation assay above, it’s slow and only measures bulk autophagy. Morphological...