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We narrowed to 1,006 results for: RING

Showing: 201 - 240 of 1006 results

  1. Thoughts on the Future of Research 2015 Boston Symposium

    Type
    Blog Post
    ...As Addgene believes many of the topics discussed during the symposium will prove incredibly important to... that the main question they wanted to focus on during the symposium was “How many graduate students/post-docs...problems in academic research and potential solutions During the first couple of sessions of the symposium it...In fact it is this very idea that postdocs are acquiring training to become professors that is provided...estimates that postdocs are paid $16.15/h before fringe benefits, less even than the $21/h [on the low ...low end] that grad students are paid before fringe benefits); what they don’t make in wages, they make up...programs as the standard option for scientists by requiring graduate students to pay for part of their tuition...

  2. From our Table to Yours: An Inside Look at Lunch at Addgene

    Type
    Blog Post
    ...cooking competition featuring a selected ingredient. Anyone who wants to participate brings in a dish that .... The four of us would eat together every day, sharing ideas for the company, but also talking about sports...a culture of openness and trust that comes from caring about your colleagues on a personal level, and ...special occasions. However, employees regularly bring in baked goods and we have a few unique lunchtime... from the software folks and started Hack Days. During Hack Days, teams at Addgene will forgo their regular...website. Free lunch is provided for participants during the Hack Days, and we’ve seen great success at ...university departments to establish, with the goal of fostering better communication between labs. Scientists ...

  3. Some Like it Hot: Thermostable GeoCas9

    Type
    Blog Post
    ...naturally occurring Cas9 diversity via the Integral Microbial Genomes (IMG) database, Harrington et al discovered...are available from Addgene. Happy CRISPRing! References Harrington, Lucas, et al. "A thermostable Cas9...Yes Longevity in Serum?* No Yes Table 1. Comparing SpCas9 and GeoCas9. *For in vitro Cas9 RNP activity...temperatures much warmer than SpCas9 can tolerate. Harrington et al tested GeoCas9’s love of heat in two ways...can introduce genomic edits in mammalian cells. Harrington et al tested GeoCas9’s editing skills by transfecting...had similar levels of editing. See figure 4 in Harrington et al for more examples of GeoCas9’s editing ...sleeve is its increased longevity in human serum. Harrington et al suspected GeoCas9 might have higher stability...

  4. A Guide to Starting Your Own Journal Club

    Type
    Blog Post
    ... once a week for several months at a time during the spring and fall semesters. Our scheduling was in ...in this situation, considering stopping (temporarily or permanently) the recurring meeting or perhaps ...Enter: the journal club.  A journal club is a recurring meeting amongst scientists that aims to help stay... At my former academic department, we had two recurring journal clubs on Immunology and Microbial Pathogenesis...club? Where will the journal club take place? Preparing for the first meeting There are many important...Addgene bioinformatics journal club! What happens during journal club? There are many common ways to run...meeting using teleconferencing software. Any file sharing service should also serve to share the materials...

  5. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...in more genome engineering technologies? Browse others on Addgene's Genome Engineering Guide. Fire ...libraries expand upon the CRISPR family of plasmids by offering a lentivirus-based mechanism for sgRNA delivery...protein and cell ageing, as the spectrum shift occurs during Timer protein life due to a form of oxidation (...transcriptomes, as well as further optimization through engineering, Ed Boyden’s lab has identified a variety of ...field of optogenetics. Using a rational protein engineering approach, they designed Opto-RTKs, which activate...flavin cofactors and dimerize on exposure to light, bringing the intracellular kinase domains into contact ...activity levels and limited sensitivity under the stringently low expression conditions necessary to avoid ...

  6. How to Be an Excellent Trainee

    Type
    Blog Post
    ...to build a bench mentor-mentee relationship and bring your best self to the training experience.  The ... isn’t discussed often enough is the hands on mentoring you receive after you’ve found your mentor, though...lab (and possibly much more, depending on the mentoring style within the research group). The bench mentor-mentee...you don’t do proper preparation and pay attention during training, then you won’t be very proficient afterwards...how to prepare and things to keep an eye out for during training. Preparation before bench work Are there... something then you’ve done previously) you can bring those questions to your training! All of these things... the in-person training component. Documenting during training It’s hard to recall a lot of details, ...

  7. 25 Work From Home Tips for Scientists

    Type
    Blog Post
    ...purchasing a gift card to use later or ordering take-out) or volunteering to foster a pet, for example. If ...is based in the lab. If you’re unsure what to do during this period, check out these tips and resources...time for exercise. Many studios and gyms are now offering online classes and participating in these is a...class twice a week at Addgene HQ, but now we’re sharing links to online yoga classes. Get outdoors. Taking...ride is a great activity for both exercise and clearing your mind. And because the outdoors is so huge...staying away from others.  Meditate. Meditating during the day is a great way to take a mental break, ...activities like team lunches, a virtual soup lunch, and sharing photos of our pets. #SocialDistancing may send...

  8. From Lab Bench to Leadership: Transitioning into Management as a PhD

    Type
    Blog Post
    ...involves a different skill set: mentoring, setting strategic goals, and fostering team success. This shift requires...wet lab wasn’t the right path for me. I began exploring alternative career trajectories within the industry... reinvent myself and redefine the value I could bring to potential employers. Over time, I discovered ...broader vision rather than the day-to-day details. Ignoring individual differencesManaging a team isn’t “one...motivations, strengths, and learning styles. For example, during my time as a lab scientist, I worked with a diverse...frustration, micromanagement, and disengagement. By tailoring my leadership approach to each person, I was able...company by taking on responsibilities such as mentoring a junior colleague. Guiding an intern, in particular...

  9. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...studying membrane dynamics, for monitoring cargo trafficking, and for discovering organelle contact sites. This...signaling pathways to the cell cycle. Good tools for measuring intracellular pH are required in order to determine...endogenous gene activation. This system works by bringing together an anchor protein and activator protein...MCS/SacI site in pUCXKT, removing the stop codons during backbone digestion and providing the missing codons...utilizes the bulldozer-like quality of the ribosome during translation, which knocks off all mRNA binding ...NLS-MS2-RFP) the reporter mRNA, which contains the tethering sites for the fluorescent proteins and an inducible...both the GFP and the RFP proteins in the nucleus. During the first round of translation, the GFP proteins...

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

    Type
    Blog Post
    ...presentations and a handout on protocol sharing, material sharing, computational reproducibility, and tools...wet lab protocols, reagent sharing, bioinformatic tools, data and code sharing, and data analysis and visualization...identify the right opportunities, he was able to bring together talented and motivated groups and individuals...s contributions by Tyler Ford included reagent sharing and were presented at the ASPB meeting by the collaborators... exclude different modules for this workshop. Empowering others to have the materials and resources to... everything that you would want to know before, during, and after organizing such a workshop. Finally,...Finally, the group put a lot of effort into ensuring that the content of the materials and the workshops themselves...

  11. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    ...genome engineering workhorse You rarely hear the word ‘CRISPR’ without shortly thereafter hearing ‘Cas9...Cas family of proteins has many useful genome engineering enzymes, each with their own unique features ... and quirks. Here, we will review the genome engineering abilities of the lesser-known Cas enzymes while...Cas9’. The Cas9 nuclease is a genome engineering powerhouse, and for good reason. It is guided by an RNA...strand break. Cas9 has been studied for genome engineering purposes far more than any other Cas family protein...making Cas9 an adaptable choice for many genome engineering endeavors. For more information on Cas9 variants...particularly convenient for targeting experiments requiring single strand DNA generation, such as homology...

  12. How Do I Place an Addgene Order Using a PO Number?

    Type
    Blog Post
    ... and Laura O'Regan Ordering jeans online from your favorite store is easy. Ordering genes on plasmids ...to other ordering questions here. In addition to answers to common questions about the ordering process...most frequently asked questions related to the ordering process: I placed an order through my university...will be able to enter your PO number as payment during checkout. Placing your order online has some benefits...Add all your items to your cart. Click checkout. During the payment step of checkout, you will have the... invoice to the billing address that you listed during checkout. This invoice will reference your Addgene...Addgene order number and PO number that you entered during checkout. At this time, your organization will ...

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

    Type
    Blog Post
    ...approved 91,023 during the first 3 quarters of 2017. As an international researcher and aspiring entrepreneur... has been a growing shift away from employers sponsoring individuals for green cards because of the cost...have different roles, titles, and responsibilities during their careers - being aware of this can help take...regulations define a "specialty occupation" as requiring theoretical and practical application of a body...limited to biotechnology, chemistry, architecture, engineering, mathematics, physical sciences, social sciences...business specialties, theology, and the arts, and requiring the attainment of a bachelor's degree or its equivalent...work-authorization is strictly limited to employment by the sponsoring employer. On April 18, 2017, President Trump ...

  14. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...vector into the culture medium during adeno-associated virus manufacturing. Human Gene Therapy, 21(10), ... iodixanol) using a high-speed ultracentrifuge. During the spin, the virions will form a band due to their...check out the Addgene AAV Production protocol. Titering Once you’ve produced, harvested, and purified ...titer of your viral vector solution. A variety of titering methods have been developed to determine the titers...TCID50) (Zen et al., 2004).  To choose the optimal titering assay, you’ll need to understand how different...outcome and planning.  Both ddPCR- and qPCR-based titering assays measure physical titers, meaning the concentration...has to be digested with DNase before undergoing titering, to remove residual plasmid DNA carried over from...

  15. PhD Applications After COVID

    Type
    Blog Post
    ...worry about traveling during an already stressful time, and I was more relaxed during both sets of interviews...yourself to a few members of your desired department during the application process. Many institutions recommend...in case one isn’t able to respond to your email. During my PhD application cycle, I reached out to around... and make sure you cover everything you need to during the interview. (Virtual) Campus Visits After receiving... financial and logistical obstacles could make touring potential programs and universities stressful or...Through the virtual systems created and promoted during COVID-19, online meetings with current students...worked with students to maintain these time goals during COVID-19? How are students funded? Are there any...

  16. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...genome engineering across multiple species Recent discoveries in the fields of genome engineering and synthetic...multiplex automated genome engineering (MAGE) (1). MAGE uses recombineering (2) to simultaneously incorporate...gene of interest. Although the parts involved in altering gene expression are relatively well known, particularly...labs who wish to harness this powerful genome engineering tool. Bassett, et al. J Genet Genomics. 2015...loci (multiplexing). Currently, the only genome engineering method in bacteria that enables rapid, automated...combinatorial genomic libraries. MAGE has allowed genome-engineering endeavours of unparalleled complexity in Escherichia...mutant enables transient suppression of DNA repair during oligonucleotide integration, allowing MAGE in otherwise...

  17. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    ... with limited time for one-on-one mentoring. His solution? Bringing his research into the classroom. I...textbooks” (Brian, 1996). But is it possible to bring the kinds of transformative educational experiences...dehydrogenase (MDH), a citric acid cycle enzyme he studied during graduate school. As a stable, easily purified protein...interest to other researchers in their field. Indeed, sharing results with the broader scientific community represents...all shaped his teaching style, with the former offering excellent training in improvisation and the latter...Jessica Bell, Lisa Gentile, Ellis Bell, and Amy Springer), this group now connects MDH-focused CUREs at...available technical and pedagogical resources, offers mentoring for instructors, and provides opportunities for...

  18. The Blue Flame Award: Celebrating Addgene's Most Requested Depositors

    Type
    Blog Post
    ...community. Sharing is caring We’re thrilled that our depositors reap the benefits of sharing plasmids with...hooked on sharing because, as UNC’s Bryan Roth says, Addgene has “literally enabled sharing with 100's...plasmids deposited at Addgene enhance scientific sharing worldwide but a select few are particularly special...honor the creativity, passion, and dedication to sharing epitomized by the creators of these plasmids with...appreciate our depositor’s dedication to scientific sharing, we wanted to celebrate our depositing PI’s who...deposited in Addgene!” As you can imagine, answering plasmid requests can be a time-consuming chore...Flame Award winners- we’d like to thank you for sharing your plasmids at Addgene. You are the reason we...

  19. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ...mix. During the incubation the immobilized antibody binds to the target protein, thereby tethering it to...approaches allow for optimized strategies for differing amounts of target proteins or antibody bonding...immunoprecipitation and simply refers to the process of anchoring an antibody, often called the capture antibody...eliminating the risk of the primary antibody interfering with downstream assays. This is particularly ...for an IP, samples are typically lysed in a non-denaturing buffer containing non-ionic detergents, such ...proteins, including those in the nucleus, more stringent buffers may be needed. Since many factors, including...Frequently, the antibodies used for a standard, denaturing Western blot will not be suitable for an IP, ...

  20. Revamp Your Lab Meetings With Creative Virtual Collaboration

    Type
    Blog Post
    ...team building and bring back some degrees of belonging that was probably lost during these weird times...Discussing data with colleagues can be more difficult during the pandemic era, with fewer opportunities for ...   Implement new tools for creative interaction During Zoom meetings, different tools can be used to promote...The annotate function can also make the screen-sharing experience more interactive. The button is on the...collaborative cloud tools such as Google Docs and Sheets during Zoom lab meetings to chip in ideas and take minutes.... This document can be shared and serve as a springboard for future discussion. Collaboration and creativity...great functionality that is widely used and could bring back a taste of that casual kitchen chatter, ultimately...

  21. Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer

    Type
    Blog Post
    ...immune system to fight progressing cancers. It was inspiring to participate in the 2017 Cell Symposium on Cancer... are coming together to tackle the challenge of curing cancer in cross-disciplinary projects and collaborations...reflect how cancer/immunology researchers are preparing and carrying out this multi-pronged attack. Inflammation...our recent post “Cancer and the Immune System: Deciphering the Relationship” by our guest blogger Subhadra...CRISPR/Cas9 methodology with T cell receptor engineering (Ref 5)  as described by Blue Flame Addgene depositor...Professor Michel Sadelain from the Memorial Sloan Kettering Cancer Center. In contrast to using randomly integrating..., CRISPR/Cas9 genome editing allows targeted engineering of the TCR locus. He also highlighted the UCSF...

  22. Addgene’s Viral Service - Why Virus? Why Now?

    Type
    Blog Post
    ...to all of our generous, sharing depositors. Over 3,000 laboratories are sharing plasmids (with 16 labs ...labs currently sharing viral preps) through the Addgene repository. Without these scientists and the support...to have such an impact on scientific materials sharing. I would also like to recognize a few scientists...Addgene has been successful in helping thousands of sharing scientists from laboratories all over the world...Scientist Team has been developing expertise in answering questions on virus and helping provide educational...reach scientists around the world. Our Software Engineering Team (and, added in 2018, our Product Team) continues...Maintaining our excellent service and accuracy involves caring and careful Addgenies, but also a lot of good tracking...

  23. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    ...the work by using the principles of recombineering. In recombineering, bacteria express phage recombination...REPLACR-mutagenesis, harnesses the power of bacterial recombineering to create insertions, deletions, and substitutions...how to replace your SDM method with REPLACR (Recombineering of Ends of Linearized Plasmids After PCR). ...modification by homologous recombination. In most recombineering strategies, only 3-4 phage proteins are needed...directly transformed into bacteria expressing recombineering proteins Red/ET from a temperature-sensitive...bacteria are subsequently grown at 37 °C, the recombineering plasmid is not maintained. Recombination efficiency... the need for selection usually required in recombineering. REPLACR-mutagenesis is also well suited for...

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

    Type
    Blog Post
    ...into their scoring functions. Many tools have also opted to mix-and-match different scoring algorithms... by comparing their past results to the designs suggested by each tool. For those who are entering sgRNA...facilitate most experimental applications in CRISPR engineering. When compared to that collective utility, it... and approximate methods. The details of their scoring function can be found in their online tool’s documentation...is by far the most common with most tools also requiring that the user specify an organism. Output diversity... genome”, “large genome”, and “organism biased scoring”. Trends: Many tools can handle genomes of any ...solution but are often packaged as offline tools requiring expertise in one programming language or another...

  25. Addgene Stands with the Scientific Community

    Type
    Blog Post
    ...their biological materials during funding transitions. Enabling resource sharing: Our operating model allows...agility as a nonprofit resource can prove to be vital during this period of constrained funding. Addgene is ...lost amid lab closures or project terminations, ensuring that scientific progress made through years of...structures and indirect cost caps more efficiently. Offering free educational resources: Our eBooks, videos...productivity and efficiency with fewer resources. Fostering new partnerships: We're seeking and expanding ...strengthening our commitment to collaboration and resource sharing, we can navigate this difficult period together...what support would be most valuable from Addgene during this time. Your input will directly shape our response...

  26. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    ...consideration when titering a lentivirus is the cell line that is used for titering. Typically researchers... my (lentiviral) generation The first step in delivering genes via lentivirus is deciding what packaging... competence can arise from recombination events during viral production and poses a biosafety risk to ... plasmids in order to obtain the best titers. Requiring an additional plasmid for transfection can make... are required to generate RCL. To overcome the titering hurdles of third generation packaging systems,...will choose a relatively permissive cell line for titering in order to obtain the best titer possible. While... One should also consider the specifics of the titering transduction experiment. Media choice, reaction...

  27. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ... and morphology, rendering their continued use inadvisable for experiments requiring reproducible results...visible and you should always ensure nothing is covering it. Turning on the UV light 3-4 times a week for...remove all jewellery as well as that watch you’re wearing before you start. Also, avoid unnecessary talking...hood. Planning your experiments thoroughly before entering the hood is another great way to prevent contamination...you’ll know exactly what materials you’ll need to bring into the hood and can wipe them all down before ...air flow around your work area. Make sure to only bring required materials into the hood. This will give...sure cells don't stay out too long While you’re preparing things, make sure your cells don’t stay out too...

  28. Drew Endy Introduces the Biobrick Public Agreement Plasmid Collection

    Type
    Blog Post
    ...accomplish is to make the engineering of biology much easier. So you could imagine engineering the entirety of ...Stanford develops engineered DNA systems capable of storing data and computing inside living cells. He's also...an organization working to advance biological engineering openly so as to benefit all people and the planet... what we think of as an operating system for engineering an entire cell. Typically when people use plasmids...cell. Until we get a handle on precise, forward engineering of the central dogma at the genome scale, it'...collection of genetic parts for precisely forward-engineering the central dogma across many genes simultaneously...been 40 years since the invention of genetic engineering. What's incredible is the bioeconomy is already...

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

    Type
    Blog Post
    ... generated will enter the culture medium during subculturing processes or from the air if the cell line...that are generated (while talking or pipetting) during working in the laminar airflow are also among the...source of the problem is the media, used for subculturing, which is available in both liquid and powdered... before sampling (Uphoff and Drexler, 2011). Culturing on agar plates/broth is considered the ‘gold standard...separate lab should be made available for cell culturing and maintenance of the cell lines. Providing such...coats. Have separate footwear for the lab to avoid bringing in environmental contaminants.  Figure...Addgenie works in the tissue culture hood while wearing PPE. Photo by Kate Harten DeMaio. Cell lines...

  30. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    ... cell or neuron type may not work well for a neighboring cell type, even within the same brain region ...What AAV vector is best for my specific system?  Sharing data and community concerns AAV vectors are complex...particular importance to these efforts is the open sharing of negative data.  Negative AAV data is highly ...why they don’t share, many express concern about sharing negative data they cannot explain, perhaps out ...therefore actively work to remove the stigma around sharing negative data and instead focus on getting this...if many scientists choose to help each other by sharing their data. So take advantage of it and get your...Lukner, H., Kogermann, K., & Sepp, T. (2021). Data sharing practices and data availability upon request differ...

  31. Addgene is Expanding Our Viral Vector Service!

    Type
    Blog Post
    ... January 2025.  What will the ordering process look like?  The ordering process is split into two steps...   Figure 1: The process of requesting and ordering a Packaged on Request viral vector prep.  ... research and expanded choice when it comes to ordering high-quality materials from the Addgene repository...Retrograde (AAVrg) capsids.  Please be aware that this ordering process will look slightly different from our ...steps, requesting and ordering. This split allows us to provide researchers with increased choice and flexibility...flexibility; while ensuring we can maintain high standards for both our depositors and requestors.  Requesting.... There is no charge for the request process.  Ordering If your request is approved, you will receive ...

  32. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...without the undesirable side products generated during chemical synthesis of these molecules. To make ...recombinant protein purification and metabolic engineering. If instead you want to build genetic circuits...biology MoClo applications, but there are more! Engineering disease resistance through gene stacking, increasing...kit has a focus on transformation and genome engineering. This is a specialized system with parts including...Multiplex MoClo Toolkit for Extensive and Flexible Engineering of Saccharomyces Cerevisiae.” ACS Synthetic Biology...Mitochondria.” BMC Biotechnology, vol. 17, no. 1, Springer Science and Business Media LLC, 13 Nov. 2017. ...Vector Toolkit and Parts Library for Advanced Engineering of Plant Genomes.” The Plant Genome, vol. 16,...

  33. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ... she's either day-dreaming about exploring sci-fi worlds or exploring nature in the real world. Additional...ICE on Synthego’s blog. CRISPR-based genome engineering has revolutionized the gene editing field by ...edits performed over multiple experiments and comparing the robustness, accuracy, and speed with existing... of the sample name. If there was a minor error during processing, the window shows a yellow checked circle...with non-wild type sequence) as determined by comparing the edited trace to the control trace. In the ...Value - When the ICE linear regression is computed during generation of the ICE Score, the Pearson correlation...sizes in the entire edited population of genomes. Hovering over each bar of the indel plot shows the size...

  34. An Inside Look into Addgene's Dev Team

    Type
    Blog Post
    ... Figure 1: Addgene’s Agile Approach Sharing is caring There are many moving parts to our software...overall soundness of the software design. Each of us brings slightly different cognitive biases as a result...defined chunks, called stories, and work on stories during short periods of time called sprints. The team ...different technical approaches that may come up during discussion.  One example of agility at work is ...we recently reduced the number of steps involved during checkout. Demo details for this would be something...diversifying our own knowledge of the software, sharing knowledge allows for life to happen without impacting...developing new features and fixing bugs: we take time during each sprint to do some maintenance work on our ...

  35. Addgene at the ASM Microbe 2016 Conference

    Type
    Blog Post
    ...Wisconsin lab for a summer during my PhD and is responsible for my enduring love of cheese curds. We’...long line for bag check was definitely worth it! Hearing about his passion for global health and the various...one of the highlights of the conference. It was inspiring to see how microbiology research is improving ...researchers who weren’t aware of Addgene’s plasmid sharing services, as well as #happydepositors who stopped...leverage their expertise as we advance our mission of sharing science. I got to check out many of the posters...about our mission of open science and resource sharing and was also a great place to catch up with former...distribution service! Did you see a must-have plasmid during ASM? Let us know what’s missing in the collection...

  36. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...combines CRISPR and recombineering-based MAGE (Multiplex Automated Genome Engineering) technology. pMA7CR... a backbone-targeting gRNA cassette for plasmid curing through "self-destruction" after induction with...CRMAGE is much more efficient than traditional recombineering, with 96-99% efficiency for point mutations...for a single round of editing, and a subsequent curing protocol that requires only 2-3 hours incubation...References 1. Maddalo, Danilo, et al. “In vivo engineering of oncogenic chromosomal rearrangements with ..., et al. “Multiplex CRISPR/Cas9-based genome engineering from a single lentiviral vector.” Nucleic Acids.... Sakuma, Tetsushi, et al. “Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector...

  37. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...repair and genome engineering CRISPR-Cas9 has revolutionized the genome engineering world and made targeted...Originally written by Marcy Patrick and Mary Gearing on Mar 12, 2015; updated by Christina Mork, Jul ... fill in the resulting ssDNA. A single HJ forms during this process and resolves by cleavage of the crossed...immediately applicable in DSB-induced genome engineering, it has biological importance for the repair ...intact. In other words, Cas9 will keep cutting and repairing your target DNA until a mutation is introduced...Homologous Recombination. Jasin M and Rothstein R. Cold Spring Harb Perspect Biol (2013). PubMed. DNA double-strand... (2018). Pubmed. CRISPR References: Genome engineering using the CRISPR-Cas9 system. Ran FA, et al. ...

  38. Finding and Joining Your Dream Lab

    Type
    Blog Post
    ...other to be physically present at certain times during the day? We asked three lab heads how they go about... If you are looking to join a lab, you may be wondering “What is this PI looking for in a new researcher...of CVs collected from throughout the year since hiring opportunities arise at various times. If the timing...will provide a solid foundation on which to thrive during time spent as a graduate student or postdoc. Although...chosen research field is important, the end goal during this time is to be trained to think and execute.... Or perhaps, the lab has a reputation for not entering collaborations. Alternatively, perhaps the lab...to ask lab heads, lab members and lab neighbors during the interview process. What if I want a non-academic...

  39. Is this the right place for me? 8 tactics for choosing a lab

    Type
    Blog Post
    ...some of these questions in the sections below. Mentoring relationships can help identify your priorities...Alumni meetings can be the most informative way of gathering data. Former lab members can give open, revealing... Know who's boss You sign on to a lab for the mentoring only to find out that the lab head is an absentee...You are not allowed to speak to lab members alone during the interview period. People seem to be hiding ...about discussing data. There’s hesitation before answering questions about lab culture. Support for diverse...ensure that you are expanding your options and preparing for both academic and non-academic career paths...evidence of industry collaboration? If you are considering a career at a small, teaching college you might...

  40. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    ...methods for physical shearing exist as well, such as sonication and needle shearing. Essentially, any method... with the lab during its recent move to the University of Georgia). She enjoys pondering how phages get...between 1 and 3 kb results in optimal likelihood of capturing a whole RBP gene without causing issues associated...following colony PCR of 10-20 colonies using primers bordering the multiple cloning site of the vector. In order...saturate each with lysis solution (~3 ml each), pouring off any excess. When your protein induction is .... Leaving the membrane overnight in the freezer during one of these steps is also a convenient stopping...through the day, spending time with her horse, and wondering what her cat is thinking. Find her on Twitter ...
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