...cutters. You can also sort enzymes by clicking on column heads. Clicking on a particular restriction enzyme...plasmid maps!" After thoughtful design, vetting, and tweaking, we’re excited to announce that our plasmid and...GSL Biotech's SnapGene Server Software. With the backing of SnapGene’s sequence viewer software and extensive...identities of most features labeled directly on them making them much easier to quickly identify. Improved ...With this more functional display, you can start thinking about your next cloning experiment early on. Beyond...Maps This is the first tab you’ll land on after clicking the “Analyze Sequence” button. The maps hosted...similar hover over features. As with the map tab, clicking on two separate locations in the linear sequence...

... and then condensing that and making it accessible to students.” Making science approachable is often ...This involves going in front of a camera and kind of walking them through the steps of, for example, cutting...And I’m like, 'Well, I’m here so let’s do it!'" Working as a subject matter expert Caitlin started out ...job as an SME: “I write curriculum for students taking science courses online, which is a blast. I have...an introduction that gives a basic overview of kinematics in physics, to developing a lab about mitosis...eScience Labs as part of their curriculum by either taking existing curriculum and customizing it for schools...courses encompass what they want to teach.” Besides working with students and faculty, Caitlin also collaborates...

...basic one, making sure that you have a good fundamental understanding of what you’re working on is important...years. We each have a unique “origin story,” but taking the first step into the world of academic research...glassware with brightly colored hazardous liquids. Working in a lab is nothing like a typical part-time or... and applied. The current pandemic serves as a striking example of the applicability and importance of...be unsure, it helps to have a vague sense of what kind of research you would be excited about. I know quite...do what they do. I would also highly recommend talking to other undergraduate researchers, as well as ... yourself into is really important.  As you’re talking to people, though, try to be conscious of who has...

...thawed and used to produce a set of ‘working’ stocks of cells. Working stocks should be frozen in multiple... generally ensure cell line identity and purity making it easier for comparable studies to be performed...to check their identity. For example, perhaps a working cell bank has been made by a research institution... good recordkeeping, you need to have a way of tracking how many passages the cells have undergone and... any of the Master cell vials until all of the working vials have been used up. This ensures you can retain...means you can regularly discard the cells you are working with because you have plenty of maintained freezer...freezer stocks. It is vital that the Master and Working stocks of cell lines have the passage number recorded...

...available for tracking certain cell types. Another useful tool for marking and tracking cells is the the...tools allow one to study the kinetics of cell migration and movement by marking cells at a specific time.... Pathogens - Kamenyeva et al, 2015. Cellular trafficking studies enabled by fluorescent proteins The optimization...tissues, including lymph nodes, brain, liver, gut and skin. Two-photon intravital microscopy. The video above...that some are more reliable and more efficient at tracking cells. Good two-photon probes when using a standard... cells and follicular dendritic cells within the skin and lymph node respectively. The permanent labelling...photoactivable or photoconvertible proteins, as it enables tracking throughout the lifetime of the cell.     The past...

...qualify what kind of “plate” you mean. Without context, it’s difficult to know what kind of plate they...professionals from other backgrounds at conferences or networking events. And if you’re a wet lab scientist who...collaborative lab work. Corralling context and junking jargon I suspected I knew at least one source of...they were talking about during the meetings. And “plate” certainly wasn’t the only problematic word. How...We took steps towards solving this problem by 1) making sure we qualified these words when we discussed...and racks meant we had fewer errors in our LIMS tracking system. Though you might not have to communicate...creating shared experimental protocols. Clarify: Asking for clarification during a meeting can help get...

...release IL-36 cytokines. Keratinocytes are the main source of IL-36 cytokines in the skin making it important...the uncontrolled activation of the interleukin-36 (IL-36) cytokine family resulting in the activation ...disease research. This year is our fourth year working with the Rare Genomics Institute by providing plasmid-based...pustular psoriasis is a chronic autoinflammatory skin disease estimated to affect 0.27 to 4.6 per 10,000...surrounding IL-36 biology. “Little is known how IL-36 cytokines actually get out of keratinocytes and induce their...researcher at University Hospital Zurich. IL-36 cytokines are a potent inducer of an autoinflammatory loop... our biosensor repository as we find ourselves lacking several crucial biosensors that would allow us ...

... worth taking time to honestly evaluate your own style of work and communication before making this important...attention you need? Are you good at asking for help? If you are good at asking for and getting help, size might... toss data-driven reasoning out the window when making decisions. We think scientists make choices based...based on emotions and assumptions. I’m not just talking about simple things like “Which route should I ...members can give you a fuller picture. Consider talking to people in the lab across the hall or in the ...prematurely? Do lab alumni stay in the same field taking projects with them and remaining close colleagues...t choose a mentor that sees you only as a paper making machine. Choose one that is a true mentor, supporting...

...yourself. Instead of thinking “What’s my angle on attacking this problem?”, try thinking “Who can I assemble...researchers do learn MANY transferable skills while working in the lab and by offering advice on areas where...type of team can exist indefinitely. If you’re looking for this type of team in academia, look no further...an imaging core or tissue culture core, you are looking at a process-oriented team serving a particular...you find that there is not someone (or someones) taking on these responsibilities on a team to which you...is certainly necessary, but equally important is making sure all team members understand what roles they... none of that matters. If you are on a team, any kind of team, and you have nagging doubts that someone...

...of my job], so making it a success feels like a real accomplishment.” He started working on them shortly...not sure what his next step should be. He debated taking a position as an application support/sales representative... to me. She was leaving a writing position and looking for someone who might take over. I like writing... a science writer these days. I do some public speaking and a bit of video work for webinars (get tips...shortly after joining IDT when his boss was looking for someone to take over hosting them. Since then, webinars... writing group I am part of is responsible for checking the scientific accuracy of everything that goes...analysis.” Han’s biggest piece of advice for those looking to break into the field of science communications...

...back out when you are looking to change positions in the future. Anyone looking for work in today’s job...researchers do learn MANY transferable skills while working in the lab and by offering advice on areas where...student or postdoc forever, and you need to be thinking about what comes next. To that end, one of the...for your career is to expose yourself to other working professionals, and one way to do this is to attend...and connections are how you get jobs! Job fairs Speaking of jobs, it seems like an obviously important ...conference, job fairs bring you face-to-face with working professionals, with the added benefit that most...larger meetings offering resources for students seeking employment within industry or academia; although...

...recognize their MANY transferable skills learned while working in the lab and offering advice on areas where you...using creative problem-solving in the lab   Thinking Outside the Box: Individual Approaches Whether...something tangible and applicable to your situation.   Thinking Outside the Box: Team Approaches At some point...same problem. If your lab/company/team is still working towards a culture of encouraging dissent, the next...nine enemies that stand in the way of creative thinking. I won’t go through all nine here, but I’ll discuss...longest tenure, but don’t let that stop you from speaking up.  If your ideas are not well received, understand...work stalls. We become so fixated with the idea of making something the best it can be that it seems impossible...

...cells. These cells produce inflammatory cytokines - interleukin (IL)-12 and interferon gamma (IFNγ) - and...help escape immuno-surveillance, and curb cytokine and chemokine production in the tumor microenvironment...their microenvironment by producing C-C motif chemokine 22 (CCL22) and TGFβ. T-regs are suppressive immune...tolerance beyond direct proximity to the tumor; chemokine-aided activation of APCs produces modified dendritic...cancer. Surgical removal causes increases in tumor cytokines like TGFβ and IL-6. However, when strengthened...pathway, VEGF production, and CTLA4 and CD protein blocking. Other molecular targets under research include...response. Antibodies targeting the tumor-augmenting cytokines produced in the tumor microenvironment TGFβ, IL...

...addition, it’s really asking a lot of someone to be comfortable on camera while thinking about things like...Science Video Production Specialist, I’m constantly thinking about ways to improve our productions to deliver...we developed a great plan to start consistently knocking out high-quality videos for you all. The next ...products or methods, using metrics and feedback, taking a look at what works and what doesn’t, and trying...Design-Build-Test-Learn cycle adapted for video making.   Cycle One: The Ironing Board Control Center...Center With the pandemic in full swing and everyone working from home, how were we going to film anything? ... prove to be difficult. Shipping the gear, and walking everyone through the recording process was going...

...discovered that many of my old suitemates stopped talking to each other if I was not there to arbitrate for...lead to hurt feelings, low morale, poor decision making, and stalled projects. You want to identify and...experiments to address a reviewer’s concerns, you are making your hypothesis stronger. When you share data at...miscommunications in the future. Assumptions: Making incorrect assumptions can be a direct cause or ...or consequence of lapsed communication. Making some assumptions about how something should be communicated...know that a person or team should be involved in making a decision and you see no record of their input... an eye out for the phrase “I assumed they were taking care of that”, because that can indicate confusion...

...affinity for glutamate but slower kinetics; v857 has faster kinetics but lower affinity) and with a variety...with the end of year approaching. We’re always looking for something to brighten our day, whether it’s... Glutamate indicators with improved activation kinetics and localization for imaging synaptic transmission...microscopy. The resulting JEDI-2P has faster response kinetics and increased brightness, sensitivity, and photostability...pYtags illuminate RTK signaling Receptor tyrosine kinases (RTKs) are a major class of signaling receptors...spatiotemporal measurements of receptor tyrosine kinase signaling in living cells. bioRxiv 2022.08.13.503850... al. 2022). iGECInano displays better response kinetics and photochemical characteristics than previous...

...wise to start working on these skills. Neuroscientists use lots of cool tools While walking around the exhibition...discuss their VR helmets, gait tracking technologies, and brain monitors. Walking around the poster presentation...Neuroscience covers many subfields When I first began looking into the details of SfN17, I expected that almost... mathematical know-how for analysis. If you’re looking to get into neuroscience and you don’t have a background...neuroscience but are afraid you don’t have the right kind of biological expertise to get into the field, there...experiments. This helped reinforce our goals of making more AAVs and educational content to help researchers... suggest a plasmid to us, request that we start making the AAV, or help us produce new educational content...

...collision that’s taking place between CRISPR technology and synthetic biology, the tools he’s making available.... Addgene: Is this what you are working on now? Voigt: We are working on this now, we have ideas, but ...Massachusetts Institute of Technology has been “working on new experimental and theoretical methods to ...large as natural regulatory networks. Addgene: What kinds of things could we do as genetic circuits become... been putting together very large toolboxes and making them available. We’ll also be submitting parts ...objective is to give people the complete toolbox for taking on very large genetic engineering projects. Addgene...you want. The last few years, we’ve really been making tons of parts and characterizing the software to...

...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...

...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...

...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...

...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 ...

...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...

...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...

... 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...

...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...

...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...

...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...

...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...

... 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...

... 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...

...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...

...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 ...

...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...

... 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...

...’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...

...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...

...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...

...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...

...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...

...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...

...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...

...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...

...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...

...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...

...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...

...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,...

...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...

... 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...

...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...