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  1. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

    Type
    Blog Post
    ...and foreign nationals working in the life sciences community obtain their working visas, permanent resident...national researchers and entrepreneurs who are looking to continue their careers in the United States....we get elected to Congress, we are tasked with working with US immigration in its current state. As of...discussed in this post benefit from the applicant thinking about the process early, keeping good records,...trained scientist. If the candidate is ultimately looking for a role in a different functional area, then...received sponsorship) then he or she can start working in October of the same year. The Industry H-1B ...of US government grants that the researcher is working under in the lab or has won, awards, media reports...

  2. 6 Tips for Grant Writing

    Type
    Blog Post
    ...useful advice about what reviewers in your area are looking for. Remember though, you need to leave them time...The two most important things reviewers will be looking for in your research proposal are originality and...provide different insight into an area - not just working within the established network of your peers. Furthermore...techniques and interact with new people. If you are looking to move to a different university, perhaps in a...re staying in the same lab where you have been working for the last few years, you need to be very clear...Building Your Scientific Network? Check out Our Networking for Scientists Series 4. Become a science communicator...communicator More and more fellowship applications are asking for a 200-300 word explanation of your research...

  3. Mouse Modeling, Part 1: Genetically Engineered Mice

    Type
    Blog Post
    ...expression. Transgenic and knockin mice As the name suggests, transgenic and knockin mice contain an additional...Conditional knockouts or knockins A subset of transgenic mice, conditional knockout/knockin mice, use the Cre-lox...terminology you should be familiar with before working with mouse models, several common techniques used...construct incorporated randomly into the genome, knockin mice have the gene construct inserted at a specific...DNA randomly into the mouse genome. In contrast, knockin strains are generated via homologous recombination...breaks, such as ZFN and TALEN, can used to engineer knockin mice. Co-injection of a targeting vector containing...based systems have been developed for generating knockin mice but because CRISPR/Cas9 engineering favors...

  4. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...before you start. Also, avoid unnecessary talking because talking generates microbe-laden aerosols that may...that can be used as your working stock. This way you can go through your working stock vial by vial before.... UV light exposure is also harmful for eyes and skin so keep the light off when the hood is in use. Wipe...wiped with 70% ethanol immediately. Finally, avoid sticking pipettes into solution bottles directly, rather...familiar with the morphology of the cell line you’re working with so that cross-contamination can be identified...indicates that the cells have grown enough, are making their surrounding media acidic through the secretion... their original condition and can replace your working stock if it gets too old or contaminated. This ...

  5. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    ...between scientific achievements. I've found cooking, gardening, hiking, and photography to be very satisfying...and running very early on. Once your system is working, you should maximize data production with the end...strengthen your new model?  PFs need to be regularly thinking about manuscripts. The other items will follow...even a relatively simple experiment. However, by taking this time, you can help ensure that you understand... feel like it's too much work, they don't like speaking, they don't feel like they get useful feedback...you give your talk or poster to enough different kinds of audiences and if you pay attention to how the...more fundamental reason to give presentations - networking! People get to know you through your presentations...

  6. Finding Your Science Policy Path

    Type
    Blog Post
    ...and unknown. In fact, in less than two years of walking away from the bench, I have worked for seven different...the science policy community is the best part of working in this profession. The folks who work in science... mechanisms in place to ensure your effective networking---and they’re more accessible than ever thanks...trajectory. If there’s one thing everyone loves, it’s talking about themselves--so don’t be shy and reach out... toward solving one problem. It often requires thinking about a problem in a new way through the integration... in a government setting: drafting legislation, making rules, and implementing the policy within the jurisdiction...to start your science policy journey.  In fact, working in science policy doesn’t have to be a complete...

  7. Embracing Serendipity: A Crucial Element in the PhD Journey

    Type
    Blog Post
    ... a diverse range of perspectives and sparking innovative thinking that may benefit your current project...In the pursuit of knowledge and groundbreaking discoveries, the world of academia often emphasizes structured...deeply into your chosen field, don't shy away from looking beyond its confines. The beauty of serendipity ...Practical approaches Engaging in mindful meditation, taking field trips, and spending time outdoors in nature.... These unrelated ideas might hold the key to unlocking new perspectives or even revolutionizing your ...that is adaptable, creative, and poised for groundbreaking discoveries. So, on your academic journey, ... the unexpected — it might just be the key to unlocking the next frontier of knowledge. Matteo Tardelli...

  8. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

    Type
    Blog Post
    ...to a few hundred worms, making the detection of rare events difficult. Dickinson et al. sought to create...markers easy to remove after injection, they added flanking loxP sites and a heat-shock inducible Cre to generate...Figure 1: Self-excising cassette schematic from Dickinson et al. (2015). Used with permission from the Genetics...for the targeted gene. After initial testing, Dickinson et al. tested this system across 7 genes in parallel...modification. Figure 2: Schematic from Dickinson et al. (2015) illustrating the steps to fluorescently...Society of America. Applications of this system Dickinson et al. have developed a modular system that requires...with a Self-Excising Drug Selection Cassette. Dickinson DJ, Pani AM, Heppert JK, Higgins CD, Goldstein...

  9. Extracurricular Activities for a Strong Science Career Path

    Type
    Blog Post
    ...Regulatory knowledge Networking Blogging Writing Communication Networking Publicity ...space in academic research labs has more students looking for other options. While graduate programs provide...Introduce new perspectives in science and engineering Looking at your research from a broader viewpoint may expose...multiple research studies highlight the importance of taking a break and incorporating EAs into your routine...their required qualifications and skill sets Taking the time to identify and research 2 – 3 potential... a profession sounds interesting, TAKE ACTION! Taking action: Examples of extracurricular activities ...Communication Teamwork Management Leadership Networking University technology transfer office positions...

  10. Synthetic Photobiology: Optogenetics for E. coli

    Type
    Blog Post
    ...Tabor have been working on making E. coli light responsive since 2005, when Tabor was working in Voigt's lab...systems continue to be optimized by Tabor’s lab, making light-controlled gene expression in E. coli easier...two-component systems (TCSs). TCSs consist of a histidine kinase that phosphorylates a response regulator (RR). ...from cyanobacteria to a common E. coli histidine kinase. This hybrid construct senses light, and an obligate... two-component system (TCS). A hybrid histidine kinase/photosensor senses various light inputs, and an...plasmids, with one RR (ompR) encoded chromosomally. Leakiness was also an issue; some promoter activity persisted...with other pathways. They also reduced system leakiness and increased dynamic range to around 100-fold...

  11. Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility

    Type
    Blog Post
    ...such as BenchSci and FPbase are working hard to enable researchers by making it easier to find information...manuscript. Luckily, many organizations have been working to provide tools that make it easier to share in-depth...quality controlled plasmids and viral vectors. When working with a plasmid from Addgene, you can be confident... about the unreliability of antibodies. IPI is working to address this issue directly by creating well...Electronic lab notebooks also play a role here by making it easier to retain searchable, thorough notes ...techniques, an important component of reproducibility is making sure that others can understand your analyses and...results that conflict with our original hypotheses. Looking for novelty in our stories, we are additionally...

  12. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...are. Here are the questions you should be asking. What kind of control? On, off, tunable, and/or inducible...off protein expression, you are almost certainly working with an endogenous locus. The most straightforward...excision. Essentially, LoxP sites are knocked in flanking a gene of interest (or a single exon) and everything...which is more difficult than a simple knock out. Looking for some permanent knock out plasmids? Check out...suppression of expression is faster and easier than making a knockout and is a great way to temporarily reduce...advantages, such as tissue-specific targeting. Looking to permanently induce expression? Check out our...Destabilizing domains Destabilizing domains (DD) are akin to degron tags, just with a flipped mechanism. When...

  13. Celebrating 15 Years of Scientific Sharing

    Type
    Blog Post
    ... value in depositing and they are increasingly seeking us out on their own to share reagents. I love the...scientists starting their own labs get excited about making their first deposit to Addgene. We are also ...universal resource for information and data. Speaking of the website, we now have a UI/UX team. Last... any more!  Our product and software teams are working in lock step to maximize the services and features... for their dedication, flexibility, innovative thinking, and openness to team work.   Company culture... via connections and even via Twitter. We love working with local organizations like Just-a-Start, the...quality, low meeting, and low email culture. I like working here and I hear from the Addgenies that they do...

  14. Summer SciComm Series: Cognitive Load

    Type
    Blog Post
    ... to exactly what you're asking of your audience and when things aren't working out, adjust the ask.  I...: “cognitive load refers to the used amount of working memory resources. There are three types of cognitive... provide detailed evidence of knowledge new to mankind. And you probably understand how much faster it...read this paper than it would be for an undergrad taking their first upper-level class. Even so, it’s quite...more you use, the higher cognitive load you are asking your audience to take on, just to understand you...If there’s a teacher, mentor, or other educator working with your audience, ask! They are experts in what...fly.  Once you can think about everything you’re asking your learners to do, it becomes a lot easier to...

  15. Science communication: 9 Strategies to Get Your Foot in the Door

    Type
    Blog Post
    ...you a science student or early career researcher looking to break into science communication? Everybody ... festivals and outreach If you are comfortable talking in public, volunteer at science festivals. Or engage...education Do you aim to reach young people? Try networking with your local education community. Get involved...environmental groups, charities and NGOs are often looking for outreach volunteers as well. 4) Blog - Hone...’t you record a podcast or put some effort into making a video? Use your unique talent to communicate ...Association allow you to spend two to six weeks working at media outlets such as the Guardian, BBC Breakfast...education Some people get into science communication by taking a media or journalism course. Courses can help ...

  16. PhD Applications After COVID

    Type
    Blog Post
    ...programs is almost as daunting as the program itself. Picking a mentor comes with many challenges, and the flood...nosy question! The Application Process Broadly speaking, the PhD application process tends to go something... to potential mentors?” “Where do I start with picking universities?” and “How does COVID-19 impact all...program is the best fit for you and your goals. Making a List of Schools Creating a list of schools is...chance to get to know the faculty that you might be working with for the duration of your program and ask any...information about the campus even if you can’t visit. By talking with people at the institution, you can get a better...into what matters to you. It can also be nerve wracking to ask questions in front of other prospective...

  17. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

    Type
    Blog Post
    ... projects and they end up making 30 constructs. They’ve never done any kind of cloning and they don’t ...tools you are working on for plants? Patron: Our lab and others at the Sainsbury are working on a lot of...general. We are funded by a charity and since we are making all the tools for plant research here and validating...problem. MoClo is really a modern toolkit for those working in plants, including many things people already...Addgene: Do you have any particular tips for those looking to get started with MoClo? Patron: The parts will... that you need from the kit and getting started making your own constructs with Golden Gate MoClo. The...network here. She talked about the advantages of talking to people and representing yourself as a real person...

  18. Transferable Skills Guide: Identifying Your Transferable Skills

    Type
    Blog Post
    ...’ve gained experience working with similar protocols, and look - they’re seeking someone who has, to a... a specific field. While searching for jobs and making connections, avoid falling into an identity trap...retrieved your lucky socks, persevered through a nerve wracking commute complete with a disabled train, and now...this job was made for you. They must have been looking at your resume while they wrote the job description...side of an issue. Maybe you also showed critical thinking skills by dissecting their problem and piecing...organizational skills. If you are responsible for working with lots of students, managing inventories in ...pretty much any other workplace. To get your mind working, take a look at the short list of transferrable...

  19. Transferable Skills Guide: External Collaborations

    Type
    Blog Post
    ...because you are not (yet) asking for a job or a big favor - you are generally asking for some advice and guidance...researchers do learn MANY transferable skills while working in the lab and by offering advice on areas where...many methods and machines one team can learn so linking together expert teams can push a scientific project...graduate studies at Karolinska Institutet I started working with other groups. First within my department where...meet collaborators  for lunch, make a point of talking to them at a seminars, or even set up regular meetings...much more effective than simply sending a message asking how things are going. Science careers where the...studies in regard to reaching out to new people, checking in regularly, and maintaining useful notes that...

  20. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ...authors’ compared nicking the top strand first followed by the bottom strand versus nicking the bottom strand...position. The “sloppy” PCR reaction is created by making some or all of the following modifications: 1) ...packaged into phage and then extracted. Besides taking a couple days to generate the phage-derived uracil...ready to be tested in a functional screen of your liking. This method is similar to site-directed mutagenesis...with a twist since it relies on the sequential nicking, degradation, and PCR-generation of a mutant version...Supplementary Note 1 in Wrenbeck et al for details on how nicking was optimized. How should I design my mutagenic...gene of interest has a BbvCI site(s). If so, the nicking mutagenesis protocol will still work if the orientation...

  21. How to Negotiate a Successful Lab Start Up Budget

    Type
    Blog Post
    ...step by step instructions on creating a budget, making decisions based on that budget, and negotiating...timeline Start with your thesis in mind. Imagine the kind of data your dream discovery will need, and how ...should reflect salaries in the region you’ll be working. At this point, your budget bottom line costs may...position. This is important to ensure that you’re looking at institutes that have the means to support your...values. All too often, we see early investigators making unnecessary short term sacrifices that hurt their...dependent on a pathology microtome/cryostat, then making a careful decision on the condition and availability... and limitations. Everything is negotiable. Try asking for discounts, collaborations, access to internal...

  22. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    ...sgRNA design is not possible without relying on some kind of pre-analysis. At the end of 2014, I began developing...contents spill to the ground. There is something akin to a frenzy as the crowd tries to weigh and determine...each item changes from what the reviewers so painstakingly assessed to something a bit more representative...community as a whole has yet to settle on any true kind of value. I’ve dubbed this the “Piñata Effect”, ...question is then, should we use all tools as some kind of a meta/Frankenstein app? Or perhaps there is ...2013 about 11 tools have been added every year.   Breaking down the barriers: My view on current sgRNA design...released with advanced functionality, it seems these kinds of features are delegated to other more suitable...

  23. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    ...affinity and slower kinetics iGluSnFR3 v857: decreased affinity and faster kinetics which are illustrated... it is selectively activated by glutamate, and tweaking it to improve its responsiveness, they showed ...Improve sensor’s photophysical properties such as kinetics, ∆F/F0, SNR, dynamic range, photostability, quantum...v82) and lime green (v857) variants differing in kinetics, affinity, and ∆F/F0. Introduce the two variants..., GPI, and SGZ, to improve neuronal membrane trafficking and localization. Improved photophysical properties...al…, Supplementary Figure 2.   Improved trafficking and localization through different C-terminal...“Glutamate indicators with improved activation kinetics and localiation for imaging synaptic transmission...

  24. Four Factors that Differentiate the Stem Cell Field

    Type
    Blog Post
    ...Scientists are known for asking "What? and "How?" , but they are not always known for asking "Should we?". Scientists...discussed his work with Parkinson's Disease; Hideyuki Okano presented ground-breaking work on spinal cord ...primary Addgene contact for many years, I've been looking forward to seeing how the sharing of his OSKM factors...clinical trials for ALS.   I have an uncle with Parkinson's, a half-brother who is a quadriplegic due to ...techniques are neither fast nor inexpensive.  After talking to numerous poster presenters, it was gratifying... Eric J. Perkins...

  25. Addgene Welcomes John Doench to the Advisory Board

    Type
    Blog Post
    ...last for several days.” Working with CRISPR Recently Doench has been working on developing CRISPR pooled...not, well, your grade is lower.' ... When you are waking up on a Saturday at like 7 am to go check your ...” Doench has spent most of his research career working on miRNAs and RNAi. He first heard about RNAi at...technology development alongside it.” Doench starting working with David Root in the then RNAi Platform, which...investigate the human genome. Doench says he enjoys working on technology development under the Platform model... but work in true collaboration with scientists asking biological questions. Doench loves that he still... “very much in line with the overall mission of making science easier and more transparent. That’s largely...

  26. Finding and Joining Your Dream Lab

    Type
    Blog Post
    ...consider when seeking your dream lab. The principal investigator's point of view If you are looking to join...decision. A science trainee will spend 4-7 years working for one person and with a group who all strive ...join a lab, you may be wondering “What is this PI looking for in a new researcher?,” “How do I convince the...typically go through a rotation, which reveals their working habits, personality, and gives them the opportunity...style to spur you to succeed or you might enjoy working closely with others to accomplish your goals. You... You must understand your own style before making this choice.  How big is the lab? While larger labs ... Dr. George Church (Harvard Medical School) for taking the time to speak with us about adding new scientists...

  27. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...protein-peptide locking, Mark Howarth’s lab has developed a new tool for peptide-peptide locking. The new technology...light-activated receptor tyrosine kinases Receptor tyrosine kinases (RTKs) are a large class of cell-surface...informational videos, download protocols, and more! Looking for background information on CRISPR technology...Easy-to-use tools to monitor these features were lacking until Roberta Gottlieb and Zhen Yan labs developed...described as “knocksideways” (a British idiom for “taking by surprise”) by Margaret Robinson’s lab, and more...mitochondrial membrane. Sequestration is accomplished by knocking down your endogenous protein of interest and co-expressing... blue and green light-excitable and has faster kinetics than those of other channelrhodopsins. CnChR1,...

  28. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    ...improve resolution, use a stacking gel on top of the resolving gel. A stacking gel typically has a different...buffer, blocking the membrane  from binding to the antibodies during the stain. The ideal blocking buffer...frequently include suggestions for working concentrations, blocking buffers, and incubation times. If ...interested in making your own knockout line for validation, please view this tutorial from Stuart Orkin's and Daniel...of the proteins in a sample migrate through the stacking gel at the same pace and enter the resolving gel...

  29. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    ...by your library. After blocking for 1 h at room temperature with gentle shaking, gently wick off excess...display little to no homology to anything else, making it difficult to quickly identify possible gene ... overexpression in E. coli, such as pET30a. If working with DNA that is refractory to enzymatic manipulation...the T7 polymerase and a few proteases knocked out making them particularly useful for protein production...harvested dose of cells (preferably in log phase) in blocking solution (around 108 CFU/mL is a good place to...start) and add your membranes to the solution, shaking gently for 1 h at room temperature. This will allow...onto nitrocellulose and carry out the steps from blocking the membrane to probing with cells in order to...

  30. Q&A with Karen Katz and Carl Paratore, the Newest Members of the Addgene Board of Directors

    Type
    Blog Post
    ...information, making it available to everyone, that creates more breakthrough and innovation. With any kind of open...'s operations. We’re looking for those “what could go wrong scenarios” and making sure we have adequate...than just picking up the phone and having a discussion. By only relying on email and never picking up the... more about Addgene and heard that Addgene was looking for new board members. Paratore: Kristen Darby,...goals were and so I decided to join. I’ve been working with Addgene since 2014 so I’ve been able to watch...

  31. Why and how to start studying chromatin (with Addgene's help)

    Type
    Blog Post
    ... from Leah Bury of Microscopic Art. I started thinking about this problem when I recently attended the...the transcription of a gene a megabase away, or looking at the histone methylation status at a double-strand...double-strand break that needs repairing. They may be looking at chromatin at the level of nucleosomes, loops... other areas of biological research. If you’re looking for resources that will help you study how chromatin...with Addgene, but you’ll find many great tools by clicking on the links below and visiting their associated...Sharon Dent Archa Fox Elaine Fuchs Kai Ge Robert Kingston Thoru Pederson Phil Sharp Julie Ahringer Alexander... listed here. If you don’t see the gene you’re looking for on this list, please consider searching directly...

  32. Reproducibility for Everyone: Lessons from an Open Science Collaboration

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    Blog Post
    ...organizations with similar ideas who were also thinking about reproducibility and shared similar desires...Fortunately, the eLife ambassadors group was already thinking about ways to tackle irreproducibility. Benjamin...organizational skills. Meanwhile, Addgene was also thinking about ways to address irreproducibility and thanks... post-workshop surveys, recording results, and tracking material downloads. This was particularly helpful...community. Lessons learned Instead of continuously talking about a problem and potential solutions, take actionable..., but how do you get them to connect? Through networking of course! Thanks to individuals in the group...right time. Be inclusive and seek solutions by working with communities, not for them - Another strength...

  33. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    ...aerosols that are generated (while talking or pipetting) during working in the laminar airflow are also ...providing proper training and guidance to the people working in the lab facility. This can be done by training...talk, laugh, sneeze or cough. Wear gloves while working and discard them after use. Human contact can spread... spread contamination when you are working without gloves or masks. Make sure to change the gloves and...inside the laminar airflow before and while you are working. Perform routine checks for mycoplasma contamination... should be cleaned regularly and dampness of any kind should be avoided. Distilled water provided to maintain...because you are not the first person to face this kind of problem. Educate others when you can and help...

  34. Design Tips for Prime Editing

    Type
    Blog Post
    ...tool of choice. Nicking sgRNAs PE3 and PE5: These tools use an sgRNA to guide nicking of the unedited...editors bound to sgRNAs only drive nicking (right). Start with nicking sgRNAs ~50 bp from the prime editing... Figure 7: Nicking sgRNAs can be designed to target edited DNA, so that nicking only occurs after... prevents the Cas9 nickase from re-binding and nicking the newly synthesized strand before heteroduplex...BioRender.com. PE3b and PE5b: In this approach, the nicking sgRNA is designed to bind only after the edit is...

  35. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    ... to worm motion. Working on the Cat Microbiome Project Another project we are working with is the cat ...computer scientists, engineers, and biologists working together on a variety of projects including the...cat microbiome project. This is a project looking at the makeup of the bacterial species in the gut of the...expensive to make it practical - but we have members looking at how to re-use it as a platform for monitoring...makerspace somewhere near you. The place to start looking for your local space is the Hackerspace website...Victoria. After a decade at Microsoft Research working on speech recognition, he returned to school to...manufacturing tools – a 3D printer, a laser cutter, woodworking tools, and a metalshop. After two summers in ...

  36. The Scientific Conference Poster Session: Tips for Success

    Type
    Blog Post
    ...provide constructive criticism, and ask thought-provoking questions. For example, if you’re a virologist...help build your presentation. For example, when thinking about discussing a figure, jot down the question...poster where the presenter spends the entire time talking to the poster, as opposed to engaging with you?...Even the most prepared of us can find ourselves talking to our posters, so if you find yourself in this...let them know while you appreciate the thought-provoking question, you are unable to answer it at this ...give the “elevator pitch,” of his/her project – kindly let the presenter know you are very interested ...interested in viewing which has drawn a large crowd, making it difficult to see the actual poster or interact...

  37. Experimenting with New Careers while in Grad School

    Type
    Blog Post
    ...while still working towards your degree. For me, experimenting with the career and networking opportunities...workshops and networking events. I took advantage of as many of these resources as I could. Networking and learning... I liked – well, I was used to experiments not working, so just like in lab, I sucked it up and moved ... like a great way to share some of the research taking place at our university with the community. I even...and David Shiffman @WhySharksMatter and see what kind of content they share and produce and how they interact...wanted to do. A few weeks into volunteering, I was talking to a little girl about fossils. After we talked...

  38. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ...very small homology regions (5-25 bp) for repair, making it easier to construct targeting vectors. Addgene...copies the sequence from a repair template with flanking sequence homology for error-free DSB repair. NHEJ...MMEJ uses regions with 5-25 bp of microhomology flanking a DSB to repair DNA. The DNA ends are chewed back...figure above, short (5-25 bp) regions of homology flanking a double-stranded break yield precise deletion... is likely to introduce additional nucleotides flanking the insertion. If the DNA ends anneal incorrectly...5’ and 3’ microhomology to the insertion locus flanking the GFP-Puro cassette. Three double stranded breaks...sets of microhomologies (5’ and 3’) to anneal, knocking the GFP-Puro cassette into the locus (see figure...

  39. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    ... unbiased potential to find things you are not looking for, but there are also benefits gained by lowering...differentially expressed genes can be the next stage in making sense of such data. These targeted screens can ...ligand (such as when a component of a microbe or a cytokine is added to a cell). With a custom CRISPR screen...example, one could study the cellular response to a cytokine known to cause the up-regulation of an important...cell). In this screen, known components of the cytokine signaling pathway could be targeted. Cells failing...the transferred cells is followed. A benefit of working with surface proteins is also that such proteins...eleven published libraries are embedded. These were kindly donated by six different research constellations...

  40. Fueled by Coffee at #SfN14

    Type
    Blog Post
    ..., D.C., we must be making a dent in the global coffee supply… My back-of-the-napkin calculations put the...attendees. And for those of you presenting, working, and networking at SfN, remember there are only 2 days ...community pulls off such an inspiring, thought-provoking, collaborative event – all while keeping the party...CRISPR-Cas tools into neuroscience experiments. Speaking of CRISPRs, we were excited to have Feng Zhang...we've spoken with are excited and passionate about making such tools open-access. Though it is time for me...

  41. Course-Based Undergraduate Research Experiences (CUREs)

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    Blog Post
    ...knowledge, shaping their trainees’ very way of thinking too. Albert Einstein might have put it best when... (Brian, 1996). But is it possible to bring the kinds of transformative educational experiences afforded...happened in the lab, and he wanted to provide the same kinds of opportunities for his students. But he also found...research capabilities and laboratory and critical thinking skills (Knutson et al., 2010a,b). Classroom-based...student apprehensions about “failure” (protocols not working, hypotheses being rejected, etc.), while reminding...excellent training in improvisation and the latter making him a more effective motivator (“be like Tony Robbins...at other institutions. Provost believes these networking experiences increase student enthusiasm, and ...

  42. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    ...by (as the name suggests) cross-linking cellular proteins. Cross-linking is a chemical reaction that covalently...solutions.  Along with preserving your sample, cross-linking can block the epitope of some targets, preventing...antibody from binding (Fig.1). The level of cross-linking that occurs depends on the incubation time and ...may be able to recover good antibody binding by tweaking those parameters, but in some cases your antibody... good for epitopes that are sensitive to cross-linking, but it doesn’t preserve sample structure quite...decide which ones to use? Here are some tips for making those decisions: Always always always look at ...recovering epitope availability following cross-linking fixation; they require an additional step generally...

  43. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    ...scanned to verify that it is the correct sample. Tracking samples - it's all about location, location, location...facility). The third tube is used to fulfill orders. Breaking the (bar)code At each step in the receiving, transforming... traced. If you’re just starting a lab, or are looking for a way to organize samples within your lab, ...in house if necessary.   Tips for barcoding and tracking samples 1. Start with the tube - Barcoded tubes... information in a small space and is ideal for tracking large collections. Many companies, such as Thermo...Although we’ve developed our own computer system for tracking samples, it can be done easily with a barcode ...multiple people or even multiple labs. Careful tracking prevents us Addgenies from wasting hours of valuable...

  44. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    ...growing quickly. Taking what I loved from each of these platforms (such as social networking, visual appeal... This post was contributed by Ryan Watkins, Professor at George Washington University and developer of...average video abstract view counts.” For researchers looking to garner more attention for their work, a video...discuss in-progress research, proposed research looking for funding, or are by researchers for whom English...English is a second or third language (making publishing their research all the more challenging even when...and let the world know about your science! Ryan Watkins is a professor at George Washington University ...the Addgene Blog 6 Tools & Tips: Online Social Networking for Scientists Read about the Evolution of Scientific...

  45. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    ...Harvard, Johns Hopkins, (most of) MIT and the University of Colorado are doing, and are checking with Yale ... salaried workers receive overtime payment for working more than 40 hours per week will increase from ...scientists within academia, most particularly postdocs working in the U.S., whose current salaries are below the...scientists are paid above this threshold or for tracking the hours that these scientists work and paying...Department of Labor issued a Notice of Proposed Rulemaking, soliciting feedback by Sept 4th.  They proposed...across academia). Increased postdoc wages, overtime tracking? What are institutions doing? Institutions are...what their institutions, or institutions they're thinking about applying to, are doing - for example, your...

  46. Healthcare Consulting: A Door to the Business of Life Sciences

    Type
    Blog Post
    ... most, get pleasure from taking up such complex challenges, quickly breaking them up into smaller achievable...clients. In this realm, there are a few different kinds of challenges that consultants get called upon to...healthcare consultant, I have worked on most of these kinds of projects. Almost all of my projects, have involved... The fast paced nature of projects often means working long hours both at home and at the office. Consulting...firms are increasingly aware of the toll that long working hours and the unpredictability of schedules can... career in healthcare consulting, I found that talking to alumni, friends, and friends of friends who ... an engaging way to learn about the trends and workings of the healthcare industry. Finally, on-campus...

  47. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    ... paper into an electronic environment and hence making it searchable and shareable, but some ELNs provide...community were to: (1) prevent vendor lock in by making it easy to get data out of the ELN; and (2) facilitate...repositories. Below I describe how each of these kinds of connectivity facilitates reproducibility. Connectivity...references the record of research is incomplete, making it difficult or impossible to reproduce. As the...the graphic depicts, RSpace supports linking to all of the most commonly used online file storage tools...Slack conversations and import them into RSpace. The kind of informal conversations about research that can... are crucial to understanding research and the thinking and workflows that went into it. The ability to...

  48. An Inside Look into Addgene's Dev Team

    Type
    Blog Post
    ...Addgenies, development includes everything from tracking plasmids from the moment a deposit kit is sent...we are highly collaborative, we love teamwork. Working collaboratively allows our team to best serve Addgene... example of agility at work is our process for working on a new feature. We start with a story that describes...best approach it in our code.  Sometimes while thinking through these steps, we discover we need additional...otherwise, this isn’t how we work. We all take turns working on different parts of the code. I think the best...we might have an EMTA theme and we will all be working in the EMTA area of the code. By having everyone...Modern Greek to be easily accessible today. Roughly speaking, we translated our software from Python 2.7 to...

  49. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...similar to that in samples of interest. If you are working with cells that are amenable to genetic knockout...knockdown approach instead. In this approach, you’re looking for an antibody that detects the target in the ...can therefore be helpful to try all three when looking to label an intracellular target.   Figure...background staining. This can include using: different blocking reagents; titration of antibody concentration ...flow cytometry through this approach. If you’re looking for a flow antibody, the YCharOS (Antibody Characterization...there is a cell treatment (e.g. PMA or a specific cytokine) known to either induce or suppress the expression... which may not be available to every scientist looking to do a flow cytometry experiment. But validation...

  50. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...know exactly what you are looking for and how many of that thing you are looking for, you are ready to begin..., it’s helpful to know how many copies you are looking for. Secondly, categorize your edit – is it any...that you won’t need to screen hundreds of clones looking for your edit of interest. For all of the screening...population. This way you can ensure you aren’t looking for a needle in a haystack before going on to generate...do. A popular screening method for knockouts is Tracking of Indels by Decomposition (TIDE) (Brinkman et... there is at least ~200 base pairs of sequence flanking the editing site on either side of the amplicon... software very similar to TIDE known as TIDER (Tracking of insertions, deletions, and recombination events...
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