... communicate effectively is part of what makes working in a team productive. Take all the opportunities...conferences on interviewing, resume writing, and networking Practice your English if you need to, go to ESL... a content-useful website, write a blog Pursue speaking invitations by inviting others (be on a speaker...“learn the business” Attend events with local networking and trade organizations Another excellent perspective...People Wish They’d Learned Sooner by Peggy Klaus Looking for a mentor or want to start a peer mentoring ...

...offers all of those same advantages to researchers working in plant model systems. “The cloning is all done...trade Lohmann does have one special tip for anyone looking to try this at home in their own lab: be very careful...along at 99 percent efficiency, simply stopped working. They finally traced the problem back to the endonuclease...in the process. “We had several motivations for making GreenGate available through Addgene,” Lohmann said...system, the more building blocks will be available, making it easier for us to do our work.” References 1...

...you are studying the effect of a new tau protein kinase inhibitor on slowing diseases progression in an...group of mice, the experimental group, with the new kinase inhibitor and a second group, the positive control...control group, with an alternative but well-studied kinase known to prevent tau hyperphosphorylation. You ...experimental protocol but does not receive either protein kinase inhibitor. Since the negative control group does...variability. If one were studying the effects of tau kinase inhibitors on mice, for example, it would be better...noise. This can be done either by modifying the blocking conditions or by substituting for a more specific...

...RNase free” and refrain from talking to friends and colleagues while working with these reagents. Other ...they are relatively small and easy to work with, making them the go-to choice for studying mammalian biology...technique is still in need of improvement. Also when working with CRISPR, one must beware that not all gRNAs...blog post. Finally, always remember that you are working with RNAs, which are prone to degradation by the..., Nakauchi H (2018) Intra-embryo Gene Cassette Knockin by CRISPR/Cas9-Mediated Genome Editing with Adeno-Associated...

... Something that I struggle with as I’ve been thinking about my career options is being an overthinker...overthinker. Being an overthinker can make me scared of taking risks, afraid of leaping forward but I have learned...in a life sciences career but life has a way of making unexpected decisions. As someone who was not necessarily...what my Instagram posts would look like. While working remotely was a different experience, It was helpful... to their roles at Addgene. I had a great time speaking with Quintin Marcelino who is Addgene’s Science...

...Mentor – what do I do now? What are the benefits of taking on my own Mentee? Q1. What makes a good science...participants had certainly realized the importance of seeking out mentoring as an important aspect to their ongoing...Mentor is often one that devotes time and energy to thinking about how to be a great Mentor.    Check Out Angela...on the blog “Tenure, She Wrote” is titled, “What Kind of Mentor Do I Want To Be?” In this excellent essay...the northeast, writes a thoughtful treatise on the kind of advisor/mentor she wants to be for her new lab...

...Anterograde transport is typically used for the trafficking of organelles, such as mitochondria, as well ... dynein, while anterograde transport relies on kinesin. Different rates of speed for various forms of ...particles will infect neurons at different ratios, making these neurons fluoresce in different colors. An... The underlying mechanisms of trans-synaptic trafficking are not fully understood and may be a form of...B.T., Maguire, C.A. and Ramirez S.H. (2017). Trafficking of adenoassociated virus vectors across a model...PubMed PMID: 1381034. Watakabe, A., Ohtsuka, M., Kinoshita, M., Takaj, M.I., Isa, K., Mizukami, H., Ozawa... Resources on the Addgene Blog Retrograde AAV: Making the journey from axon to nucleus Rabies and neuronal...

...Matt Takvorian When COVID-19 researchers started working with us to help them share their reagents, we knew...COVID-19 resources are now available to scientists working in a for-profit setting.  In light of the need ...and able to listen to Addgene’s new approach of working with the industry-based scientific community. Conversations...California Institute of Technology             Johns Hopkins University                           National Institute...from the Michael J. Fox Foundation are used in Parkinson’s research.  The extensive resource from the Structural...

...their careers to get advice on choosing a lab, picking your research topic, starting a lab, and more. ...skills that employers love PhD students to have: working to deadlines, writing up clear and attractive reports...reports, being good at presentations and of course, networking and collaborating.” Getting out of the lab also...out that a PhD teaches analytical and critical thinking skills that are applicable to a variety of careers...passionate about. Check out my previous post if you're looking for more information about the benefits of pursuing...

...self-excising cassette (Dickinson et al., 2015) made this step as simple as heat-shocking to induce the expression...2013), or rescue of mutant phenotypes (unc-119) (Dickinson et al., 2013). These approaches allowed effective...knock-ins were occurring 24 hours post-injection, making screening for knock-ins much more efficient. Pure...these improvements from the Mello lab resulted in strikingly high editing efficiencies, obtaining as many ...e193–e193 . https://doi.org/10.1093/nar/gkt805 Dickinson DJ, Pani AM, Heppert JK, Higgins CD, Goldstein...

...structural biologist and protein engineer currently working at Sanofi Pharmaceuticals in Cambridge, MA. When...typical middle age weight and “exercise” consisted of taking walks in the mall. I took action, joined a gym,... potential cures. We encourage any researchers working on ACVR1 or any of the pathways that impinge upon...awareness of FOP. Please consider running with us or making a donation via the link below. Please continue ...structural biologist and protein engineer currently working at Sanofi Pharmaceuticals in Cambridge, MA. His...

...about science to everyone – not just to others working in your own field. Alternative careers are now ...necessarily bad news, but it is an argument for taking the time to hone your communication skills. So,...ll know you are on the right track if they start asking you questions. Remember that people like a good...   Consider starting up your own twitter feed or talking about and sharing some science on the one you already...ocean science, they have good advice for anyone looking to make their science matter. If you aren’t ready...

...lab. A common strategy to determine the binding kinetics of a purified protein involves immobilization ...moiety, which forms dimers, may influence binding kinetics of the target ligand. Another important consideration...-glutathione. Biotin-based immobilization for kinetic analysis Proteolytic removal of the fusion protein...protein could provide a more relevant kinetic analysis, but the trick is to utilize a second immobilization...outlined here provides a basis to determine binding kinetics using a multitude of methods including surface...

...recombinant affinity reagents. Addgene is looking forward to working with the community to create a fully ...plasmid and viral services. The first step will be working with the community to build a resource of plasmids... well validated collection. I’m thrilled to be working with Addgene to bring this resource to life,” says...10.1038/520295b Bradbury ARM, Trinklein ND, Thie H, Wilkinson IC, Tandon AK, Anderson S, Bladen CL, Jones B,...

...researching, but I decided I needed a new challenge. Looking for sales and marketing jobs Like most choices ... researchers. Many biotechnology companies are looking for PhD or Master’s level scientists who have knowledge...business-like elements. At this point you may be asking yourself, "Why would you switch the focus of your...the employer was not such that he/she would be looking at where I had published, who I had published with...scientific and marketing knowledge. For most PhDs looking at careers outside of academia, sales and marketing...

... accountability that is one major advantage of working with a mentor.The responsibility of arranging meetings...that need work or growth. Here are some ideas for making your meetings productive: Discuss logistics if...reach all the goals you set, but the process of working on them is where growth happens. Mentors bring ... day. If I skip, I get a bunch of text messages asking me, “What happened? Where are you?” My gym posse...Develop coaching skills Encourage your creative thinking Enhance leadership skills Support inclusion, diversity...

...monitoring the first round of translation or by tracking the translation of a single mRNA over time. Both...translation. Single molecule continuous translation tracking TRICK allows you to see the first round of translation...translation, but it doesn’t allow continued tracking of a given mRNA because NLS-PCP-GFP is removed by ...first ribosome that translates the mRNA. To permit tracking over time, Yan et al. devised a system that labels...keeps it in a single field of view for constant tracking. You can then directly monitor multiple rounds...

...Professor at University of Barcelona. The challenge: Making courses on genetic manipulation more hands on While...meeting with some other professors, I went home thinking about how to solve this problem. I didn’t want...your strategy. I realized that similar types of thinking go into developing gene therapy strategies for...noting any gaps in their knowledge. Practically speaking the format I decided on was to have two teams ...skills like team building, collaborative critical thinking, and communication among others. They additionally...

...much of the primary data one might require when making such a decision remains spread across literature...With thousands of scientists routinely using and thinking about FPs, it would be great to have a centralized...building advanced multi-parametric search queries.  Looking for a bright, monomeric FP published in the last...suggestions. If you’re not sure exactly what you’re looking for, the Table of Fluorescent Protein Properties...that set of proteins.  For computer-savvy users looking to incorporate FPbase data in their own projects...

... Otito Izu is a PhD Candidate working under the supervision of Akindele Oluwatosin Adeyi at the University...University of Ibadan, Nigeriaunder supervision of Dr. Akindele Oluwatosin Adeyi, University of Ibadan, Nigeria...research related to space medicine. Now he aims on tracking pollution in rivers by employing synthetic biology...be able to support diverse groups of scientists working in a wide range of research fields and geographical...

...requested dozens of times in 2 weeks. This is a striking example of the contributions that scientists are...to shared materials, and then pay it forward by making the new materials they develop available to the...has been able to rapidly propagate this strain, making it available to scientists around the world. This...research by removing logistic hurdles to continue making materials available rapidly to scientists (Nature...Yuen K-Y (2020) Comparative tropism, replication kinetics, and cell damage profiling of SARS-CoV-2 and SARS-CoV...

...culture towards a more generous and rewarding way of making science.   How we started Crowdfight-COVID19...as a collaborative platform to help researchers working on COVID-19. Its origins trace back to March 2020...-19, an online platform to connect researchers working on COVID-19 with scientists of all areas who volunteered...idea behind the platform was simple: any person working on COVID-19 could file a request on our website...We are a community of researchers who believe in making science less of an individual effort and more of...

...important part of being a “good scientific citizen” and making sure you do what you can to help your fellow researchers...consistent way so other researchers can use them, to making sure that methods and protocols are available and... can be published. To a scientist, all of this checking and formatting can feel like yet another hurdle...for being transparent in their communication or making their data and reagents readily available. What...confident that everyone’s enthusiasm for actually making it happen would increase dramatically. I’m not ...

...protein AraC binds O and I1 sites upstream of pBad, blocking transcription. The addition of arabinose causes... cAMP and represses pBad, decreasing promoter leakiness.   Other examples of chemically induced promoters... be compatible with your experimental system. Leakiness If you’re expressing a gene that may be toxic,...inducible promoters are also known for their low leakiness. Inducibility level How much do you want to induce... inducible promoters can vary a lot. If you’re looking for high inducibility, the Tet-On system may be...

... in Your Working Day" video from Science Media Savvy At this point, you might be be thinking social media...first blog post out of boredom. I had just started working in a new lab and I was the only person in the lab...first post so I signed up to write a second. While working on this post, I was offered and accepted a position...on the Addgene blog Read about online social networking for scientists Learn why scientists should be...

...can leave them at room temperature when you’re working with them, but avoid freeze-thaw cycles, as they...bind to amine groups present in some conjugates, blocking the conjugation reaction. Many commercial antibodies...for your ELISA and Western, I’d highly recommend sticking with what works.  To balance this out, however...in your machine before you make your purchase. Picking a single fluorophore for an experiment is easy,...accurate read and reduce needed compensation.    Working with antibodies can get a bit complicated, but ...

...toolbox openUC2 are trying to overcome this hurdle by taking these limitations into account in the development...software (left) vs. open hardware (right).    Taking this all together, the essential ingredients of...newly developed techniques are the key enablers for making assemblies open-source. The sharing of digital ...produced quickly with the printer, act here as a kind of "teleporter," since a part developed in Germany...way of communicating in science brings stronger networking across laboratory and disciplinary boundaries...

...channel just for asking people if they can write or review things and another for asking specific science...depositors and customers are asking. So how can you get involved? Keep asking your burning questions and...review to catch those pesky typos, and hit publish. Making content is an iterative process. We are constantly...

...responsibilities. After my second postdoc, I realized that working in the wet lab wasn’t the right path for me. I ...style of a new, junior team member who had values, working styles, and skills that were quite different from...from them. I started at my previous company by taking on responsibilities such as mentoring a junior ...Beyond Academia: Stories and Strategies for PhDs Making the Leap to Industry." He successfully transitioned...Resources  Resources for aspiring managers Books: The Making of a Manager by Julie Zhuo Radical Candor by Kim...

... account, you can begin the deposit process by clicking "Submit Online Deposit" (for online submission... menu: Figure 1: Begin the deposit process by clicking "Submit Online Deposit" or "Download Deposit Spreadsheet...  Figure 4: Add plasmids to the article by clicking “Add Plasmid to this Article” (left) and entering...button and jump around to the different pages by clicking on the appropriate boxes at the top of the page...the plasmid to copy from the drop-down menu and clicking the white "Copy" button. *Note: The "Copy" button...Submit Deposit" button to complete the data entry. Clicking the "Submit Deposit" button will allow you to ...

... the mice to increase their nose-poking behavior. Strong nose-poking behavior is exactly what was seen...a channelrhodopsin, such as cell localization, kinetics, and spectral properties, must be optimized simultaneously...photocurrent strength, wavelength sensitivity, and off-kinetics.   This model was then used to predict which channelrhodopsins...photocurrent amplitude, wavelength sensitivity, and off-kinetics correlated with the model’s predictions.  Three...

...on everything, including your skin. Here are some general tips for working with RNA, even if you are using...basics of isolating RNA without a kit. Tips for working with RNA (whether you are using a kit or not) Although..., 2006).  View our protocol for RNA extraction! Making the simple protocol even more foolproof (still ...still without a kit!) As mentioned above, working with RNA requires keeping your samples cold until homogenization...

...digest/ligate methods and picking the right one can save you time if you’re working at speed or money if budget...you can explore to maximize the use of both when making plasmids. Use repositories Repositories such as...free. Plasmids ship out 2-3 days after ordering, making it a toss-up as to whether it’s faster to order...experiment. Make your work “high-throughput” Instead of making one plasmid at a time, try constructing several...

... unique cutting features and evolved PAM sites, making Cas9 an adaptable choice for many genome engineering...same mechanism as traditional spCas9. If you are looking to modify RNA, Cas9 just won’t cut it – literally...editing. Fast facts – PAM requirement: protospacer flanking sequence – A, U, or C. Best for: targeting single...use. Fast facts – PAM requirement: protospacer flanking sequence – none. Best for: targeting single stranded...single base pair edits, these enzymes are worth taking a look at. Choosing the right Cas can really put...

... it’s monumentally difficult without software. Tracking lab procedures also gives you access to hard data...repeated. We also now have a good interface to begin tracking analytics data for lab procedures. With this data...are distinct from their contents. You should be tracking both, you should easily be able to move contents...less time getting up and running and more time working on interesting biotechnology code.There are valid...reasons to choose a non-Python stack, but if you're working with scientists it's unlikely they will have exposure...

... There are currently 17 sites around the USA undertaking BEST programs, including local schools in Boston...contributing to the NIH’s Biomedical Research Workforce Working Group. Along with her participation in this panel...Pubmed Central PMCID: PMC3787295. 6. Marder, Eve. "Looking out for future scientists." eLife 3 (2014): e04901...PMC4185419. 7. NIH 2012. Biomedical research workforce working group report. http://acd.od.nih.gov/biomedical_...in Science in Our Career Posts! Get Advice on Networking at Scientific Meetings from Addgene Executive...

...edit and homology to the genomic DNA locus. After nicking by Cas9, the homologous pegRNA sequence hybridizes..., as it can edit near to or far from PAM sites, making it less constrained by PAM availability than other...previously used when they developed base editing. By nicking the non-edited strand, they can cause the cell ...efficiencies 2-3-fold. However, the process of double nicking increases indel formation slightly. Designing the...the edited strand, as in the PE3b system, guides nicking of the unedited strand only after editing has occurred... ✓ ✓ ✓   ✓ Second sgRNA to guide nicking of the unedited strand Favor the edited strand...

...understand the risks of the specific virus you’re working with before you work with it. This will help you...required in BSL-1 and required in BSL-2 labs. When working with lentivirus, many organizations require or ...can quickly be removed and disposed of without risking exposure. You can request disposable gowns from...researchers choose to wear eye protection when they are working in a biosafety cabinet, even if it is not required...used for BSL-2 waste, and of course, if you are working with a substance in a biosafety cabinet, it needs...

...include a lot more. Font size: Making your poster readable While we are talking about font, remember this:...important to reflect on these impressions when thinking about crafting your own poster. What sets a scientific...that it will display your data most prominently, making it the star of your presentation. Portrait. This...presented some standard formatting guidelines for making a scientific poster, you shouldn’t hesitate to ...

...with the NPA where I gained invaluable experience working on and later chairing the Advocacy Committee. I... I’m now furthering my leadership experience working at the Association for Research in Vision and Ophthalmology...website. By volunteering, you can gain experience working with other people mostly online and on monthly ...resulted in actionable tasks and I made a point of sticking to deadlines and performing the work I agreed ...members changed, I had to learn how to develop new working relationships with these individuals who often ...

... months have in store for us. If you’re thinking about making the switch from academia to entrepreneurship...entrepreneurs. To make things more interesting, we are working in a new technical discipline that nobody understands...few jobs available. So, we spent a lot of time talking about what we could do to make ourselves marketable...marketable to the wider world and occasionally thinking about how we could bypass the establishment hiring...

... they become confluent, taking both early- and late-arising clones, and making sure we always maintain...densities - adding to the expense of dilution cloning. Taking a page from bacterial cloning, we reasoned that...cloning rings. A picture of a 60 mm dish ready for picking is shown in figure 1.   Find Additional Useful ...firmer 2% agarose gel solution discussed above for picking. Many thanks to our guest blogger Iris Lindberg...

...Additional opportunities: Tracking intracellular pathogens In addition to tracking the location of proteins...University of Queensland) to observe membrane trafficking events in vivo. Locate your protein and see who...your images using subjective judgement by simply “looking at them”. Modern cameras and image analysis software...limited by the wavelength of light and, practically speaking, for common lab microscopes the detection limit...

...alleles. Although CRISPR is very good at making knockouts, knocking in large sections of DNA can be more ...negative selection marker like the HSV thymidine kinase (HSV-tk) is included just outside one of the homology...” the resistance gene through the insertion of flanking LoxP sequences, you can remove the gene through...described in the preceding sections are simple ways of knocking out a non-essential gene of interest. However,...

...You’ve spent days and weeks thinking of an amazing project. You’ve written your protocols, designed your...enzymes laying around the lab. If not, you’ll be looking at about $50-$60 per enzyme. Once you’ve got a ... of the restriction enzymes and overall you’re looking at a price of a few dollars per reaction for traditional...relatively new to the cloning game, however, or simply looking for a more streamlined approach, you might want...inserts: There are some cloning methods that make working with multiple inserts a breeze. Among them are ...

...a bench scientist would looking at a genome by eye. We came up with a ranking formula that does just this...In the rest of this article, we'll explain the thinking behind the tool, its key features, how it works...information from Crispresso plus a summary per batch. Ranking guides for HDR in-depth In other tools, guides ... engineer with the Chan Zuckerberg Initiative, working on assignment at the Biohub in San Francisco. Before...

...broken down into different types, with Cas3 systems making up type I, the most abundant type in the Class ...consistent expression can still be a challenge. A multitasking Cas Most Cas effectors have one job — Cas see...away at both the non-target and target strands by making cuts every so often. Cutting both strands ensures...carried out much like the name implies — after nicking, Cas3 reels in the non-target strand of DNA much... for Cas3’s biggest trick. Cas3 likes to bluff, making you into think it’s more powerful than it is. The...

...Despite these grim statistics, new strategies for attacking cancer are yielding promising results in the laboratory...universally-dividing behavior of cancer cells, attacking the cellular machinery necessary for division....in the field of cancer research has focused on picking apart the complex cellular pathways involved in...found across all TCGA tumor samples is represented, making the results of any such study as clinically relevant...Addgene's cancer collection pages Scientists are making progress. New resources and technologies become...

...co-founder of the BioBricks Foundation, an organization working to advance biological engineering openly so as ...thing at a time. What the BioBricks Foundation is seeking to accomplish is to make the engineering of biology...is in reference to George Boole who in 1854 was thinking about logic and analyzing human language and observed... be made freely available? Endy: When we start thinking about languages, humans have two types of languages...really plugging into the "stuff sector" if you will, making chemicals and materials, and we anticipate plugging...

...using the application correctly is imperative. Picking your chemistry  The two most common qPCR chemistries...thermodynamic and experimental parameters (neighbor stacking energies, loop entropy effects, cation concentrations...type (such as here, here, or here).  If you’re working with samples that have not been documented in the... plateau stage after exponential amplification, making it difficult to properly analyze your data.  To...experiment! The model assumes the amplification kinetics of the target and control genes are fairly equal... e-book on applying MIQE guidelines! With these kinds of resources, you will soon become a qPCR master...