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We narrowed to 931 results for: TIM

Showing: 241 - 270 of 931 results
  1. Plasmids 101: Choosing an Antibiotic Resistance Gene

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    Blog Post
    ...can be shortened since the plated cells will have time to produce beta-lactamase prior to division, instead...effects of the antibiotic. This buys you a little more time, since bacteria won’t divide immediately after it...beta-lactamase. Pros: Widely available, cost-effective, timesaving for transformations. Cons: Prone to satellite... Pros: Very stable, interchangeable with Amp, timesaving for transformations. Con: Expensive.   Kanamycin...resistance Choosing the best antibiotic resistance ultimately comes down to your applications and needs. The...selecting!   Antibiotic Resistance Stability Time Saving?  Works for multiple organisms? Compatible...storage temperature and pH on the stability of antimicrobial agents in MIC trays. Journal of Clinical Microbiology...
  2. AAVs in Retinal Gene Therapy

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    Blog Post
    ...some genetic diseases. I didn’t know it at the time, but gene therapy had fallen - plummeted really ... organ that can be monitored non-invasively over time. Importantly, it is an immune privileged compartment...destructive inflammatory immune response. At the time, adenoviral vectors were the most commonly used ...cells - AAV - were becoming available. At the same time advances in human genetics led to the identification...brought all these key elements together at the right time and place. But it is the hard work and persistence...deliver via AAV.  Dual vectors strategies are being optimized to overcome this limitation, with limited success...designing expression cassettes to regulate and optimize transgene levels, and large scale manufacturing...
  3. New Tools Enable CRISPRa for Neuroscience Applications

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    Blog Post
    ...neuroscience, including transgenic mice, neuron-optimized viral vectors, and high-throughput screening approaches...Figure 1b), and sgRNAs can be found here. Neuron-optimized viral vectors Alternatively, Cas9 activators can...recently by the Day Lab, who developed neuron-optimized lentiviral vectors for in vitro and in vivo CRISPRa...with BioRender.com. The Day Lab used their optimized vectors to highlight a key advantage of CRISPRa... to neuronal physiology. The Day Lab’s neuron-optimized lentiviral vectors can be found here. High-throughput...not all neuroscience applications require such optimized tools. This is elegantly demonstrated by the Qi...PMC6214761. Savell, Katherine E., et al. "A neuron-optimized CRISPR/dCas9 activation system for robust and ...
  4. Designing Your Chalk Talk for the Academic Job Interview

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    Blog Post
    ...your screen result may or may not actually help. Ultimately, if you cannot describe the likely outcome with...grant. Either way, think realistically about the time and resources required to feasibly accomplish the... address the aims in more depth in the remaining time. The typical chalk talk structure is: What is the... details. Make the most of your limited speaking time. Beyond the information you seek to convey in your...feedback. It’s also helpful to practice at least a few times with people that are outside your lab and outside...or while the committee is arriving. Spend as much time as possible facing the committee and not the whiteboard...secretory proteins in the Endoplasmic Reticulum, optimization of fluorescent proteins,  and live cell imaging...
  5. Using Video to Share Your Science: We Share Science

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    Blog Post
    ...us, such as finding time to read all of these articles. Most of us hardly have time to keep up with all... our own topic. Only on occasion do we then have time to read about research from other disciplines, or... of interdisciplinary research while at the same time increasing interdisciplinary citations in their ...videos of 3 to 5 minutes provide more than enough time for researchers to describe their science, and for... Rely on Circadian Rhythm and Melatonin for the Timing of Nocturnal Courtship Vocalization. The video ...youtu.be/EHHhiawIdWM Saving salmon, one embryo at a time – A video by LeeAnne Thayer, a Ph.D. candidate in...
  6. Tips for CRISPR Gene Editing in Mice

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    Blog Post
    ... gene you’d like to manipulate in mind. Now it’s time to consider where you would like to attack this ..., 2014. Not all gRNA’s are effective. At the time of writing many of the specifics surrounding why... remain unknown, but fret not! The key to saving time and energy during your in vivo work is to screen...similar plasmids for gRNA expression), one gRNA at a time. After 48 hours of incubation, you assess the level...technique, you can assess several sgRNA’s at the same time, and choose the most effective ones to increase ...validated your gRNAs for use in mammalian cells, it’s time to use them in mouse embryos. When researchers first...mouse embryos to develop. Once they are born, it’s time to screen them for your desired edit. Good news!...
  7. Getting the Most from Your Lentiviral Transduction

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    Blog Post
    ..., in actuality, many find using lentivirus to be time consuming, difficult, and lacking in reproducibility...enough to promote cell growth. Overgrown cultures stimulate quiescence, the resting phase of the cell cycle...efficiencies. Cultures should only be passaged 20-30 times before discarding and thawing a fresh vial. In many...prefer to collect and pool the harvests from multiple time points while others opt for a single harvest after...lentivirus are determined by the envelop protein. Many times lentiviral vectors are engineered with non-native...such variables as cell density, volume, incubation time, and MOI can all impact the success of your lentiviral... S., Guo, S., Xu, J., Kappes, J.C., Hel, Z. “Optimization of the transductional efficiency of lentiviral...
  8. Data Freedom: The Expansion of Data Sharing in Research Publications

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    Blog Post
    ...is that access of this data is to be made at the time of manuscript submission. The rationale includes...these were those from researchers who wanted more time to mine their own data, with some smaller labs feeling... originator was oblivious to, no matter how much time they had. It could be argued that data generators...having exclusive access to data and enjoy reasonable time to make sense of it, with their decision to publish... specific (expensive) software that evolves over time such that prior formats are no longer supported/... especially for young researchers who may be intimidated by having to contact more senior colleagues. ... kinases implicated in human diseases. His spare time is spent trying to raise the alpha isoform of GSK...
  9. Michael Koeris' Journey from Grad Student to Entrepreneur: The Story of Sample 6

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    Blog Post
    ...this time, Dr. Koeris and Professor Timothy Lu (a graduate student in the Collins' lab at the time) got.... As you'll hear Dr. Koeris explain however, the time was not right for the development of phage-based...listening! Download the podcast here! Podcast breakdown (Time : Topic) 0:00 - 1:25 : Introduction 1:25 - 6:19 ...   We'd like to thank Dr. Koeris for taking the time to talk to us and taking up a position on the Addgene...
  10. The Advances Behind The World's First Personalized CRISPR Treatment

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    Blog Post
    ...transplant — creating intense time pressure for the research team. Ultimately, the lifesaving effort was ...contribute to more in the future. Figure 1: A timeline of CRISPR base editing advances that led to KJ...indel mutations that plagued CRISPR editing at the time. KJ was born with a C→T mutation on one allele of... of off-target editing and longer genome search times. Using a machine learning approach, the lab proposed...patients. While these gRNAs would still need to be optimized and assessed for off-target risks, such a modular...development of genetic therapies even further. In the meantime, base editing will undergo even more improvements...
  11. Using Phosphoserine to Study Protein Phosphorylation

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    Blog Post
    ...interest as possible, some optimization will be necessary. To begin, one should optimize protein production using...phosphoserine directly into recombinant proteins (5). Optimization of this system defined three components required...phosphoserine, the Sep-tRNA synthetase, and EF-Sep – an optimized elongation factor that accepts Sep-charged tRNAs... have worked most extensively worked with and optimized Sep-OTSλ and can recommend its use (6).  2. C321...that can be used as a positive control during optimization experiments. This version of GFP will contain...purification strategies; expression should be optimized for parameters such as induction temperature and...
  12. Optogenetics + CRISPR, Using Light to Control Genome Editing

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    Blog Post
    ...develop an optimized photoactivatable CRISPR gene activation system, but settled on a different optimal fusion...to use light to externally control the location, timing, and reversibility of the genome editing process...researchers tested a variety of combinations to optimize both fusion proteins. The best performing combination...fusion protein combination. The optimized LACE system consisted of: 1) CIBN-dCas9-CIBN, where CIBN is the... prevent DNA binding in the dark. Upon light stimulation, the pdDronpa moieties move apart, allowing DNA...LOV2 has a C-terminal helix that unfolds when stimulated by blue light. When fused to the catalytic site...chromatin architecture, it is useful to regulate the time in which dCas9 binding occurs. Bubeck et al. expressed...
  13. From our Table to Yours: An Inside Look at Lunch at Addgene

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    Blog Post
    ...continued this tradition. We now have over 30 full-time employees, so we created one long lunch table and...bring in baked goods and we have a few unique lunchtime traditions that might be fun to incorporate into... take a break from their everyday tasks and make time for special projects? We borrowed an idea from the... success at accomplishing “we-should-really-make-time-for-that” type projects. Cross-team “Random Lunches...between labs. Scientists are often willing to make time for free food, and these lunches would provide a... as important to take a break from work to share time with the people around us. It makes the quality ...
  14. Mapping the 4D nucleome with CRISPR/Cas9

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    Blog Post
    ...nucleome, the organization of the nucleus in space and time, and to understanding how nuclear organization varies... change in response to the cell cycle or other stimuli.   To create a colorful Cas9 toolbox, Ma et al....study is the method’s sensitivity. The authors estimate that a minimum of 150-200 fluorescent protein ...phototoxicity concerns and extending the potential imaging time for this technique. The NIH Common Fund has made...epigenetics and cellular reactions to external stimuli. It could also have major applications to cancer...imaging of genomic loci in living human cells by an optimized CRISPR/Cas system. Chen B, Gilbert LA, Cimini ...
  15. Finding Your Perfect Job After University

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    Blog Post
    ...more practical lab experience to later gain a full time position in a lab. I worked on the genetics of Wilms...of interest before you dive straight into a full-time position. For my part, I really enjoyed some aspects...the company was to fulfill contracts, my work sometimes felt rushed and the focus always seemed more on...has been rapidly growing. These factors left me optimistic that I would find a genetics related role soon...situation, communication between departments was sometimes difficult and the solution attempted was more ...solved with good management practices, but can sometimes go unnoticed at large organizations. Another downside...because you need to work independently most of the time and plan your own work schedule.  I really love ...
  16. Bricking Science: Portraying Scientific Reality Through LEGO

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    Blog Post
    ...videos is very time consuming; producing a 1-2 minute video takes an entire week of our free time. After taking...preconceived obstacles, optimistically grasp at limitless potential, and who sometimes wish they could live...about anything can disrupt your bench work and sometimes no culprit can be found…. In our lab we call this...practically universally loved (except for those rare times that we step on them with bare feet), to connect...
  17. ReFigure: Save Scientific Figures into Dashboards and Share Your Insights

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    Blog Post
    ...during this time more visible and thereby have a positive impact on science. ReFigure saves time, knowledge...continue reading for more details. How much time do you spend reading scientific literature? Very...experimental results but, obviously, this is not optimal. What if you could save the figures of interest... instantly and are editable by the author at any time. Insights contained in your ReFigure could be beneficial... Resources on the Addgene Blog Use Apps to Save Time and Track Your Experiments Use video to share your...
  18. Walking beside Academia and giving it a high five: My transition from graduate work to a position at Addgene

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    Blog Post
    ...s Lab (plasmid page here). Now I spend much less time wrangling bacteria and instead help Addgene fulfill...Addgene? So why did I join Addgene? Was I jaded by my time in academia? Did I run screaming from the lab like...Outreach Scientist where I spend a large amount of my time meeting with and talking to graduate students, post-docs...teenage years I was super shy - there was even a time when I was too nervous to ask store clerks for help...graduate work at Harvard, I recognized that I would ultimately want to go into science communication but thought...writing, teaching, presenting, analytical thinking, time management, information distillation, etc) are transferable...
  19. Using Addgene's New Viral Service for Your Research

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    Blog Post
    ... other reagents. Virus distribution came up time and time again with many researchers lamenting the viral...Instead of spending time and money producing virus from select vectors in the repository yourself, you...frustrating process that requires a large investment of time and resources for labs that don’t have experience...titering, and testing viral preps for them. At the same time, we’re developing ourselves as a knowledge base ...to be distributing new types of materials. At the time of publication we’ve already fulfilled more than...
  20. A Guide to Getting Started in Undergrad Research

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    Blog Post
    ... Working in a lab is nothing like a typical part-time or 9-to-5 job. I had to ask to find out what a “...Professor, Assistant Professor, Associate Professor Sometimes teaches graduate and/or undergraduate classes ...that not all advice is good advice, and it is ultimately up to us to decide whether we want to take it...mentor and choosing the right lab. Remember that your time is valuable! Don’t jump on the first opportunity...whether it’s something you want to dedicate your time and energy to. How can I make the most of my experience...to ask “dumb questions” and to make mistakes. Ultimately, though, scientists are all just people. I’ve...cultivate stronger relationships with my labmates. Sometimes what I thought was a dumb and very basic question...
  21. Celebrate National Book Month with Addgene

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    Blog Post
    ... or good - they're just exist and sometimes we benefit and sometimes we don't."     - Jennifer Tsang, ...imaginative, funny, bittersweet, and shocking at times."     - Jason Snair, Product Designer Sarum: The...of southern England. While the length may be intimidating, Ruthefurd’s writing will capture the attention...cross with each other and with the events of the times."     - Aliyah Weinstein, Marketing and Communications...International Finance. A book club could lead to smart, sometimes contentious, discussions. I presented the first...
  22. The MTA at Addgene

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    Blog Post
    ...individual MTAs every time they wanted to share materials, and the process was time-consuming and lengthy...often get positive feedback on “our” MTAs, which sometimes makes people think our MTAs are different than...transfer office is sent a deposit agreement. The One-Time Plasmid Deposit and Distribution Agreement establishes...when biological reagents are shared — both at what time and with whom. Tracking these connections allows...refined as new technology becomes available. Our one-time deposit agreement means we only have to work with...
  23. Management for Scientists: Delegating is Key

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    Blog Post
    ...just do it myself” or micromanage –you don’t have time for that and you won’t be as productive. For example... to, or what tasks you assign, the manager is ultimately responsible for outcomes. Be ready to support...assignment to discuss goals, expected outcomes, timelines and deadlines. Agree on a way to review project...dumb” and that it is better to ask before wasting time. Focus on results. What does success look like...delegation allows you to make the best use of your time and skills. It also ensures that other people in...delegation tips and practices can be found at the Time Doctor Blog "The Art of Delegation: Developing This...
  24. Advice for Scientists Starting a Lab

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    Blog Post
    ...the majority of his time in the lab.  As time went on and he was spending more time in his office, he gradually... offer support. “It is an exciting and emotional time!” George Church tries to recruit people who are ... and dealing with email can be half of my day sometimes.” He points out that the copious amounts of email...George Church (Harvard Medical School) for taking the time to speak with us about adding new scientists to ...
  25. 15 Hot Plasmids from 2017

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    Blog Post
    ...write about your plasmids in a future blog post. No time to read? Listen to our hot plasmids segment on the...CyOFP1, a large stokes shift, long fluorescence lifetime, and a stable monomeric form as determined by ...resonance energy transfer) and FLIM (Fluorescence Lifetime Imaging Microscopy) applications. To demonstrate...isolated and characterized: mScarlet (fluorescent lifetime of 3.9 ns, quantum yield of 0.70), mScarlet-I ...maturation in cells compared to mScarlet, fluorescent lifetime of 3.1 ns, quantum yield of 0.54) and mScarlet-H...photostability compared to mScarlet, fluorescent lifetime of 1.3 ns, quantum yield of 0.20). All three mScarlet...working with edited “pools” of cells, eliminating the time-consuming step of single-cell isolation. Agudelo...
  26. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

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    Blog Post
    ..., these methods can be labor intensive and quite time consuming.   As Cas9/gRNA activity is crucial for...need in the scientific community for efficient real-time Cas9 activity assays. Recently scientists have created..., integrated reporter for assaying CRISPR/Cas-stimulated HDR activity) The Madison lab created SRIRACCHA...ratio of RFP to GFP expressing cells provides a real-time read out of Cas9 activity.  By using the RFP:GFP...reporter systems provide efficient, rapid, and real-time read-outs of Cas9/gRNA efficiency. So if you are...integrated surrogate reporter for assaying CRISPR/Cas9-stimulated homology-directed repair. Journal of Biological...
  27. The Many Reasons for Irreproducible Research- and a Vaccine to Eradicate It

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    Blog Post
    ...rigorous studies will give false signals from time to time. False positives in original studies and false... provided in most published papers. The precise timing, order, or every combination of materials and treatments...preregistration Preregistration is the process of creating a time-stamped research plan in a public repository before...control, and you think there are four important times to measure that growth rate, you report the results...occurs during the course of the study, say a fifth time has unpredictably high rates, should be reported...
  28. Developing a cold tolerant E. coli using specialized chaperones

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    Blog Post
    ...out of their optimally suited temperature ranges and make them grow better at suboptimal ones, which could...are your regular next-door microbes which grow optimally in moderate temperatures (usually defined between...quantity can significantly reduce the operational time for a given process. These are extremely useful ...worth the trouble.  Enter ELSA: E. coli Learning Suboptimal Acclimatisation To resolve this impasse, our ...mesophilic chaperones would not function well under suboptimal conditions. This means recombinant proteins can...
  29. From Lab Bench to Leadership: Transitioning into Management as a PhD

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    Blog Post
    ...value I could bring to potential employers. Over time, I discovered that I not only enjoyed the technical...a cell culture experiment myself would take less time than the seemingly burdensome task of explaining...strengths, and learning styles. For example, during my time as a lab scientist, I worked with a diverse team...when managers fail to set expectations or provide timely feedback, leading to unproductive dynamics. I experienced... experienced this firsthand when it took me some time to adapt to the communication style of a new, junior...solid foundation in the basics of management. Invest time in understanding key principles through books, workshops...
  30. 15 Years of Addgene: The Top 15 Plasmids

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    Blog Post
    ...tools. How many times have these plasmids been shared in total? Just over 49,000 times! Their long-lasting...adventures...From moving office locations three times, to opening our UK office in 2014, to starting our...such as our second most requested plasmid of all time, pMD2.G. Find psPAX2. pMD2.G - Also from the Didier... use in human cell lines, the Zhang lab codon-optimized SpCas9. Using the BbsI restriction site, a pair... version is the 15th most popular plasmid of all time, but was found to have a point mutation in the puromycin...
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