...include a note about where you met IRL (in real life) and perhaps what you talked about (or how good the... this rule, but not that many). I’m asked a lot about requesting connections to complete strangers for...have been able to tell when some start-ups were about to be bought by seeing certain groups of people ...using social media as a scientists Read blog posts about networking ...

... data and the list goes on and on. As you think about your career (and your future in general), keep in...in this post benefit from the applicant thinking about the process early, keeping good records, and planning...writing, compliance, core facility management, collaboration management, grant administration, clinical operations... in professional publications written by others about the alien's work in the academic field Evidence ...people who have not worked with you directly or collaborated on a project where they provided you with feedback...Celano, CEO Propel Careers. Lauren is passionate about working with scientific individuals to find exciting...

...different SARS-CoV-2 spike variants. You can learn more about their research in this article. To see other plasmids... optimized. Browse the collection or learn more about it on the blog. COVID-19 research is happening fast...through plasmid sharing and more. As you’ve read above, some of these plasmids have been in the repository...Additional resources on the Addgene blog Learn more about mapping the SARS-CoV-2 - human protein-protein interaction... interaction Find tips for remote collaborations Browse all COVID-19 blog posts Resources on Addgene....

... posters can be. You have also probably thought about why certain posters resonate with you (or not). ...important to reflect on these impressions when thinking about crafting your own poster. What sets a scientific...Making your poster readable While we are talking about font, remember this: bigger is better! Your poster...gradient (as is shown in all of our illustrations above) so that the color fades to a lighter or darker ...conference attendance Read all of our blog posts about conferences ...

...Have you ever been stuck googling about an antibody that will be essential to your new research project...high-quality antibodies for IHC. Working with collaborators and support from the Brain Research Through ...extensive library of monoclonal antibodies; Addgene collaborates with these companies to create and provide general...pattern should align with what the atlases (mentioned above) and literature suggest, providing an expectation...open-source database, the mAb3D Atlas will foster collaboration, progress, and innovation by allowing scientists...

...resources on the Addgene blog Learn about RNP delivery Learn about CRISPR delivery methods for mammalian...Nanoblades pseudotyped with a mix of VSV-G and the baboon endogenous retrovirus Rless glycoprotein (BaEVRless...have lower off-target edits. Another great thing about Nanoblades? They’re easily produced in a BSL2 lab... guide Find CRISPR plasmids and resources Learn about viral production ...

... 2015 conference invited me to attend and speak about Addgene’s experience helping scientists share plasmids... with the first keynote presentation. He talked about the impact of the Human Genome project on its 25th...of the meeting was how much we still don’t know about RNA. One talk that really brought this home was ...presented by Todd Lowe from UCSC. Dr. Lowe talked about his work on the 500+ human tRNA genes and, until... Udi Qimron from Tel Aviv University then spoke about using CRISPR/cas to address the growing clinical...

...the work I agreed upon (just as I would in my laboratory work). Committee members have to work professionally...to work with many different types of people both above and below you. During my time at the NPA, there ...further opportunities opened up to me. As I spoke about my work with the NPA, I was ultimately invited to...society in my scientific field - ARVO (as mentioned above). I became an official member of the Committee and...important for postdocs and graduate students to think about their career options, gaining transferable skills...

... think much about management skills during your scientific training. I never thought about management ...open mind. First let’s clear up a misconception about being a manager: yes, being a manager means being...you should be doing : writing a grant, thinking about the next project, reaching out to other teams, expanding...buffer them from unnecessary pressure “coming from above”, which is when other skills such as negotiation...

...Until then, you can only wonder what I am talking about. How do researchers use physical models of protein...students work closely with local researchers to learn about their research while creating physical models of...model is an effective teaching tool. You can read about Jacob’s initial reaction to this model on his blog...will wake up in the middle of the night dreaming about the conformational gymnastics that this protein ...Additional Resources on the Addgene Blog Learn All about CRISPR Use CRISPR for Genome Visualization Tips ...

...all the current code on GitHub, ask us questions about it, or send us a pull request for improving it. ...) so that they can be used in the rank equation above. Cut-to-insert distance was fitted to experimental...below) so that it can be used in the rank equation above. Generally, the greater this distance between the...splits up the guide sequence.) If neither of the above are options, the algorithm silently mutates codons...Addgene blog Read our CRISPR 101 blog posts Learn about some considerations when designing gRNAs Find other...

...positive experience, I was still incredibly nervous about leaving academia. A lot of academics tend to view...which may take a longer time to acquire. Learning about drug discovery: Unless your academic lab is closely...information. Working as a team: Industry is all about teamwork. A single individual could not bring a ...someone with whom you can have an in-depth discussion about your project might be challenging. In an academic...contacts to get that referral and find out more about the company’s program. Once you’ve found a few positions...

... widely available, so you’ll only have to worry about cloning your transfer plasmid. The transfer plasmids...typical insect cell preparations, which we talk about below. Processing The viral supernatant is typically...available, so the amount of cloning required is about the same between insect and mammalian systems. Baculovirus...dead cells and cellular debris. While adding time (about three to four days), this second round provides ... can skip the first round of production, saving about a week of work. An important note is that this is...

...our tissues unimpeded. Andrew and Maria dreamed about the possibilities of a magnetoresponsive fluorescent...of improved response. Then, just when Andrew was about to suggest she deprioritize the project, Maria’s...with a strong commitment to exploration and collaboration. Andrew is a physicist with a background in ...potential applications.  Andrew is a frequent collaborator and is happy to give advice and support for ...interested in working on these questions or collaborating, the York Lab is eager to hear from you. Find...

...If you follow CRISPR research, you know all about using non-homologous end-joining (NHEJ) to make deletions...plasmids, GFP knock-in cell lines can be created in about a month and a half, without the need for complicated...more predictable than NHEJ. As seen in the figure above, short (5-25 bp) regions of homology flanking a ...Directed Repair and Non-homologous End Joining Read About Other Genome Engineering Tools like Sleeping Beauty... Additional Resources on Addgene.org Learn All about CRISPR on Our CRISPR Guide Pages Find CRISPR Plasmids...

...republished here with permission. Quilt is a collaborative database for genomics. In this article, Quilt...tri_methylation In the next section we’ll apply the above formula to a real-world experiment. We’ll start ...discovery and therapeutics. We’ll have more to say about clinical applications of CRISPR in the next section...Aneesh Karve is CTO at Quilt Data. Quilt is a collaborative database. Aneesh's research interests span machine...on Your CRISPR Background with CRISPR 101 Learn about CRISPR Delivery Methods Validate Your Genome Edit...

...brainstorming sessions.  Allow the group time to think about the problem before you meet. This holds the dual... meetings run smoother when people have thought about the issue ahead of time. Encourage dissent. The ... decisions while encouraging the group to think about alternative options. When a devil’s advocate is ...a few minutes every month dedicated to thinking about the future will improve your ability to innovate...to be able to continue our experiments. Thinking about these plans before you’ve run out of consumables...

...complete editing process. The MUCICAT system While the above systems are great for experiments that require only...serve to interrupt genes (or gene clusters) in metabolic engineering. So far, we’ve used the MUCICAT system...when compared to the industrial strain. In our metabolic engineering case study of N-acetylglucosamine ...Multiple induction systems to choose from. Ready for metabolic engineering with improved cargo dedicated to gene...other CRISPR methods for bacteria Find all articles about microbiology Resources on Addgene.org: Visit the...

...101 blog posts Learn more about the origin of replication Find blog posts about plasmid cloning Resources... plasmids within the same bacterial cell, think about whether plasmid incompatibility will affect your...this is expanding as we obtain more genomic data about plasmids (Shintani et al., 2015). Now, bioinformatics...of the microbiological world. (Want to know more about these plasmid types? Read our Plasmids 101: Environmental...

...deletions, the ability to analyze and interpret data about these types of edits is critical to the CRISPR workflow...Distribution and Traces - provides particular details about the indel profile of the edited sample. You can ...sequence base calls from the .ab1 file are also shown above each trace. The horizontal black underlined region...us know how it works for you! Read more details about the ICE tool in our bioRxiv paper. The new ICE CRISPR...guiding CRISPR researchers, she's either day-dreaming about exploring sci-fi worlds or exploring nature in the...

...a single grant. Either way, think realistically about the time and resources required to feasibly accomplish...interrupted frequently by committee members. As mentioned above, plan to give your 10 minute overview and address... list of common questions, check out this eBook about applying for faculty positions. You need to practice...program. You should be proud of it and be excited about getting the opportunity to realize your vision. ...blog posts Resources on Addgene.org Learn more about careers at Addgene Watch our career videos ...

...gene, Xw, except for a single base. If you heard about CRISPR, one question probably comes to mind: can...compatibility All of the base editors mentioned above are SpCas9 nickase-based and depend on the presence... HDR methods for rice crop improvement in the laboratory of Yinong Yang at PSU.   References 1. Komor,...The CRISPR-Cas Revolution, Cold Spring Harbor Laboratory (2018). Additional resources on the Addgene blog...Download the CRISPR 101 eBook Read our blog posts about plant biology Resources on Addgene.org Find all...

...a less tangible form: knowledge and information about the materials in our repository. We started to increase...by making it easier to find materials and learn about them. As early as 2007, we began grouping similar...professors, stopping by to say, “Everything I know about virus, I learned from Addgene,” or “I learned to...seeing our resources shared via social media, on collaborators’ websites, or as a recommendation in a Reddit...updating and expanding our website, we’ve learned about the importance of smaller, systemic changes, like...

...target metabolite creates an aptamer that will only bind to DFHBI in the presence of that metabolite (Fig...approach less sensitive in most applications. Learn About Fluorescent Biosensors Light-up aptamers (or, fluorescing...fluorophore protein. Years later, Samie Jaffrey’s laboratory took advantage of their deep knowledge of the...Tips for Using FRET in Your Experiments Learn more about GFP Controlling Protein Activity with GFP Resources...

...optimization within your experiment. Read on to learn more about the various controls that can be used for plasmid-based...: Expression Level of Gene X The results shown above are from a single experiment wherein Plasmid A (...be compared. The Empty Vector Control (mentioned above) could also serve as an important negative control...experimental cells. For example, in the experiment above, Plasmid A contains an shRNA that targets human ... times and on different days. Using our example above, we would test different preparations of Plasmid...

...time I heard about FRET during a journal club, my guitarist brain automatically thought about the raised...you would say. The student was of course talking about the now well-known FRET, aka Fluorescence (Förster...other. FRET reduced the scale of our perception about molecular interactions. What is FRET? In their ...Resonance Energy Transfer    Interesting Articles About FRET: Piston DW, Kremers GJ. Fluorescent protein...

...Homology-Directed Repair Learn about base editing for plants Read about four base editor reporters Additional... The BE3 system increased editing frequency to above 30% for a variety of targets in human cell lines... Liu lab created BE4max and AncBE4max mentioned above, they also made an adenine base editor with improved...cell therapy applications. Dual base editors What about combining the function of both editors into one?...also RNA base editors, which you can learn more about in this blog post: CRISPR 101: RNA Editing with ...

... your edit, it’s important to know a few things about your target cells and the desired edits. First, ...et al., NAR). It works just like TIDE (outlined above), except that it uses a 3rd sequencing trace file...great option, especially if the options outlined above aren’t realistic. Check Out Our Post on Sequencing...higher probability locations. If you’re worried about off-target effects, you can consider using a version...SpCas9-HF1, eSpCas9(1.1), or HypaCas9. Read more about Cas9 options in our blog post here. Dealing with...

... the Addgene blog Read more about SHERLOCK and DETECTR Learn more about plasmids available for COVID-...method based on CRISPR-Cas12. Cold Spring Harbor Laboratory. https://doi.org/10.1101/2020.02.29.971127 Rauch...SARS-CoV-2 Genetic Material. Cold Spring Harbor Laboratory. https://doi.org/10.1101/2020.04.20.052159 Zhang...research Resources on Addgene.org Get the basics about CRISPR in our CRISPR guide Browse our entire COVID...

...the time between being born and giving birth - of about 10 weeks. Adult mice reproduce as often as every...are great model organism not only for the reasons above but because they actually share many biological ...cells, yeast DNA is packaged into chromosomes and about 23% of yeast genes have a counterpart in humans ... become an important model organism. They share about 70% of their genes with humans and 85% of human ...in the popular model organisms series, and read about new systems being developed for emerging model organisms...

... this chain (blue) to infer or calculate things about earlier ones. In practice, multiple sources of error...condition 2.” By eye, the loading control appears about the same between the two conditions. But it isn’.... However, it throws both assumptions discussed above into question. Assumption #1: The loading control...searching the literature to see what is already known about your housekeeping protein in your context. If you...step from the POI blot, you don’t need to worry about the linear detection ranges of multiple targets....

...I spent about seven years working in research labs, and then pivoted to writing full-time in mid-2020...Synthetic Biology One of the best ways to learn about a new field is simply to know where to look and ...might be helpful, please reach out to me by email.  About the author Niko McCarty is the editor of Asimov ...curriculum at MIT. He also writes Codon, a blog about biotechnology.    ...

...subjected to the experimental condition. In the example above, a negative control could be a third group of mice...in the experimental protocol. In the experiment above, you could repeat the tau western blot several times...Baseline secondary signaling levels In addition to the above-mentioned safeguards, you’ll need to test the secondary...Flow cytometry In addition to the controls listed above, viability and compensation controls are recommended...technique before starting. For more information about antibodies and antibody-based experiments, check...

...can be used in the repressible manner described above, as well as the inducible manner detailed in our...necessary to have all 3 of the different binary systems above if they all behave very similarly?” The simple answer... at the same time.  The 3 binary systems listed above are orthogonal, meaning that they do not affect ...Srikrishnan. “Introduction and expression of genes for metabolic engineering applications in Saccharomyces cerevisiae...

...Blog Learn more about FRET Tips for Using FRET in Your Experiments Learn more about Fluorescent Protein...heme-containing proteins in whole animals, light at or above 600 nm wavelengths should be used. Researchers often... emits light at a tissue-penetrating wavelength above 600 nm. LumiFluors Schaub, F.X., et al. (3) designed...be excited by NanoLuc® luciferase (NLuc) and set about to create an in vivo imaging reporter by linking...

...Science with Tom. In this podcast, we learn more about Tom and pick his brain for advice on how to dive... Rap of Henrietta Lacks Students in Oakland rap about Henrietta Lacks - the African American woman whose...Please Don't Kill My Hive Beautifully filmed video about colony collapse disorder.     5. Plasmid Rap ...science standards and communicate science. Learn more about Tom on his youtube channel and at sciencewithtom.com...

...therapeutics at Novartis - listen to learn more about Crystal’s exciting career path, to gain insight ...Chapter 2: What's a lab group in industry? The collaborative nature of working in industry and how delegating...metrics for success. The importance of thinking about the big picture and not just the next experiment...resources in our Mentoring for Scientists Series Learn about Turning Your PhD into a Career with Joanne Kamens...

...this new way to learn about plasmid technologies. If you have any comments about the Hot Plasmids Podcast...with these efforts, you’ll find new ways to learn about science careers, lab protocols, and, of course, ...each one, plug in some headphones to learn more about the latest and greatest plasmids through this podcast...

...Plasmids 101 blog series Learn about the different grades of DNA  Read about the various ways to quantify...scientific communities using plasmid tools. Details about the cloning grade DNA beta project We will ship ...quantify DNA Resources on Addgene.org Learn about our other beta projects Read our Molecular Biology Reference...

...Find more blog posts about CRISPR Learn more about the history of CRISPR Read about other genome editing...Science Fiction (SciFi) writers have been curious about the capability of transforming the genetic code ... of California San Diego. She is working in collaboration with Prof. Alexis Komor and together they are...

... Its Fruit Is Sweet The Future? I knew little about adeno-associated Viral Vectors (AAVs) before starting...I’ll provide some tips that will help you think about how to get started with AAVs. Since my research ...understanding of receptor physiology and the effects of metabotropic and ionotropic influences on overall cell function... the biotechnology firm Biogen have recently collaborated to develop “AAV 3.0”, vectors that will have...serotonin in movement. Follow her on twitter @katrinaboards.   References 1. Russell, David W. "AAV vectors...

...blog Learn about recycling initiatives for the lab Subscribe to the Addgene blog Learn about other creative.... In addition to doing some of the quick things above, try joining or starting a sustainability project...campaign! MIT Green Labs has a lot of resources about this, and even has a list of biological samples ...

...How did you hear about post grad opportunities and when did you start thinking about them? 18:30 - 19:... top if you leverage your community, think hard about picking a good mentor, and begin considering career...19:54 : How to think about picking a post-doc project 19:55 - 22:40 : Improvements that can be made to ...

...learn about all the amazing research being done with plasmids. I heard an impressive talk about the use...pulls off such an inspiring, thought-provoking, collaborative event – all while keeping the party going! At...scientists we've spoken with are excited and passionate about making such tools open-access. Though it is time...

...updated AAV Data Hub to share curated information about AAV usage in research applications. Scientists around...world, are learning to navigate the changes brought about by the pandemic. As we adjust, we are reflecting...I have spent the last 9 months quietly learning about Addgene’s past and present, but the real promise...forward to working with Addgene colleagues, collaborators, partners, and the larger scientific community...

... questions about the ordering process, the Addgene Help Center is full of information about how to deposit...requires a quote in order to generate a PO, learn more about how to create a quote. How do I pay the invoice...Addgene Help Center If you have additional questions about our ordering process not addressed in this article...

... highlighted reagent repositories and other collaborative efforts in the scientific community to speed...store my plasmid in TE, Tris, or water? And what about temperature? Find the details in this blog post....dominates DNA quantification discussions, but what about the other methods to quantify DNA? Here we cover...Incompatibility by Leah Schiesow If you're thinking about hosting multiple plasmids in one cell, it might ...

... constructs that compose genetic circuits and metabolic pathways. For many MoClo systems, the individual...interested in recombinant protein purification and metabolic engineering. If instead you want to build genetic...designed for flexibility. The Fragmid kit described above works in both mammalian systems and Drosophila. ... Signaling in CAR T Cells. Cold Spring Harbor Laboratory, 17 Feb. 2024. Crossref, https://doi.org/10.1101...Emerging CRISPR Technologies. Cold Spring Harbor Laboratory, 27 Oct. 2023. Crossref, https://doi.org/10.1016...

...environment? If these are pressing issues in your laboratory, then FRET experiments are a viable biophysical...to consider these cases separately when talking about the tools that are available. Let’s consider the...that, if you make an FP fusion, you are inserting about 20-25 kDa of protein mass into your protein and ...together The FRET donors and acceptors described above can be paired together to measure a wide range of...of your protein that can lead to novel insights about its biology and its behavior. See the table below...

... to design gRNAs for all the purposes discussed above. Please see videos and text on the website for more...Output: A text file containing full information about the suggested gRNAs to use, including all information... hard to read. It´s suggested that you open the above text files, copy the content of the file, and paste...Additional Resources on the Addgene Blog Learn about CRISPR Screens How to Design Your gRNA Tips for ...CRISPR User Resources on Addgene.org Learn More About CRISPR Find CRISPR Screening Libraries Browse All...