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... 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...

...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, ...

...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...

...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...

...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...

...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...

...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...

... 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 ...

...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 ...

...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...

...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...

...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...

... 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...

...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...

...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, ...

...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...

...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...

...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...

...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...

...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...