...Assembly. Gibson Assembly allows the production of scarless DNA constructs using homologous regions to guide...the joining reaction. BioBrick Assembly will leave scar regions at the site of fragment joining, but this...use of Type IIS restriction enzymes results in scarless assembly. Finally, the NEBuilder® HiFi DNA Assembly...fragment overlap sections, while still creating scarless plasmid inserts.  Want to Learn More about DNA...reset and ligate mismatched overhangs, yielding scarless DNA assembly, and allowing fragments to be taken...used directly in DNA assembly will still yield scarless assemblies. Overview of NEBuilder HiFi DNA assembly...and primer design results in covalently joined, scarless DNA – perfect for transforma­­­tion or as a PCR...

...daughters playing with some cards and I had a quick flashback to time spent playing a card game called “Magic”...this game was played. These rules restricted which cards could be played when and, depending upon who your... 100% student engagement. I figured, an engaging card game could help me get the engagement I wanted. ...disease case. Then, each team received a whole set of cards (Promoters, Inserts, Tags, Origins of Replication...each element to use, they had to place the selected card on the board game. They had 35 minutes to develop...and some were so enthusiastic that they took the cards and the board with them. My impression was that ...

...components to convert the cargo from RNA to DNA and permanently insert the cargo into the host genome. Lentiviral-specific...purposes. The primary plasmid with the delivered cargo (often a gene) is called the transfer plasmid while... most viral experiments is to deliver a genetic cargo, typically an expression construct, to cells/organisms..., though. Viral packaging and entry The genetic cargo requires viral packaging to infiltrate and integrate...functions to infiltrate the host cell and deposit viral cargo. Gene expression While AAV often does not integrate...inverted terminal repeats (ITRs) is the promoter and cargo, in that order. This will all be ssDNA within the...

...right path for me. I began exploring alternative career trajectories within the industry, but the transition...fulfilment in supporting my team’s development and career growth. This realization led me to pursue a shift...I discovered strategies to effectively pivot my career development and align it with these new goals, ...final choice Management is not a one-way path. As careers evolve, many professionals move between technical...that managing isn’t for you at this point in your career—and to step back into a technical role. My advice...transitions as an integral part of your broader career development.   Matteo Tardelli authored the book...metabolism. He now guides fellow PhDs in their careers with actionable steps and execution plans.   Resources...

...on Addgene.org Learn more about Careers at Addgene Watch out career videos  ...addressed in numerous essays and workshops (see the Careers essays in the journals Nature and Science, for ...what will be needed to achieve your intended next career stage and create a timeline for achieving these...designing and interpreting experiments, as well as career guidance. In my own experience, great mentors often...needed, this is a strong indicator that this person cares about your science and your development. Taken together...'ll be better prepared for your future intended career and you'll become a better scientist in the process...getting a faculty position Academic v. Non-Academic Career Decisions Check out our transferable skills guide...

...the available sgRNA-cargo constructs utilize a U6 promoter and contain the RNA cargo cassette inserted ...to observe intra- and post-Golgi segregation of cargo during their transport. Finally, the system can ...MluI-XhoI-SalI) chaining results in a translatable ligation scar (Val-Glu) between each module to permit easy construction...compared to other viral systems due to its large cargo capacity, minimal cytotoxicity, and broad host-tropism...and their collaborators in Robert Campbell’s lab carried out five rounds of random mutagenesis on a library...CRISPR-Display: A novel CRISPR technology for targeting RNA cargos to specific DNA loci John Rinn’s lab is interested...CRISPR-Display uses the dCas9 enzyme to deploy large RNA cargos to specific DNA loci. In CRISPR-Display, your RNA...

...Bacterial Colonies can be found below: Career videos Ever needed career advice or wonder what it’s like to...ll hear all about their career paths, future goals, and get some useful career advice.  Our first video...you to find content relevant to your research and career goals, or to answer any questions you might have...to work at a biotech nonprofit? In our Career Video series, we feature interviews with real Addgenies ...

...transposition, genetic cargos must be flanked by the L and R ends - thus, scarless insertions are not currently...efficient and programmable insertions of large DNA cargos into bacterial genomes. This new capability was...DNA (aka mini-transposon), which contains the DNA cargo of interest flanked by ~50-150 bp transposon end... which offer simple transformations for smaller cargos, and the ability to easily cure cells of the plasmid...spacer(s) into the CRISPR array Clone desired DNA cargo into the mini-transposon Transform plasmid(s) into...programmable insertions of even very large genetic cargo, at one or multiple desired genomic loci. Many applications...

...found on the UK Career Site for Researchers called Vitae. Vitae recently published a career development framework...learned a lot. Whether you are pursuing an academic career path or moving in a nonacademic direction, there... put you in position to pursue a diverse set of career paths. Check out Joanne's Reddit AMA Here are some...defining characteristics of nonacademic science careers is that a team is almost always involved. Demonstrating... framework for career skills. Download a summary PDF here or try out the online Researcher Development...

...including biotin carboxylase and decarboxylases. These enzymes catalyze the transfer of carboxyl groups to organic...important cellular metabolites. Biotin acts as a carboxyl carrier from one compound to the other. These biotin...lysine in the entire organism on the biotin carboxyl carrier protein (BCCP). Discovery of the biotin-avidin...

...using chitosan-complexed single-walled carbon nanotube carriers. Nat Nanotechnol 14:447–455 . https://...small RNAs in their respective species. Addgene carries >30 “empty gRNA” backbones which contain a plant...ubiquitin promoters (Belhaj et al., 2013). Addgene carries Cas9-containing plasmids for knockout, activation...CRISPR-Cas9 components into plants. Nanoparticles (carbon nanotubes, Kwak et al., 2019), DNA nanostructures...addition to the plasmids described above, Addgene carries several useful CRISPR toolkits for creating plant...Targeted delivery of nanomaterials with chemical cargoes in plants enabled by a biorecognition motif. Nat...

...thinking about my career options is being an overthinker. Being an overthinker can make me scared of taking risks... no idea how to translate them into real world careers. I figured this internship would be a perfect way...was certain I would not end up in a life sciences career but life has a way of making unexpected decisions...future. They made me rethink having a structured career because our mindsets and needs can change. Having...say with certainty that Addgene is a company that cares - not only in terms of its employees but in general...

...pAAV-Ef1a-DIO mScarlet AAV5 Controls Karl Deisseroth New serotype pAAV_hSyn-PdCO-mScarlet-WPRE AAV5 Optogenetics... a whole host of (awesome) research not easily carried out before. We have a bunch of other new AAV vectors...AAV-GfaABC1D-Cre-4x6T AAV5 Recombinases Stanley Thomas Carmichael New viral prep pAAV-EF1a-fDIO-Cre AAV5 Recombinases...Ofer Yizhar New viral prep pAAV_hSyn-DIO-PdCO-mScarlet-WPRE AAV1, AAV5 Optogenetics Ofer Yizhar New...Edward Boyden New serotype pAAV-Ef1a-DIO-ChRmine-mScarlet-WPRE AAV1 Optogenetics Karl Deisseroth New serotype...

...including a Christmas Chemistry symposium and a career night. The society also visits high schools to ...become involved with. The society provides talks and career support for a range of different scientific disciplines...talks for both students and the general public, career events, and networking meetups. The society hopes...interested in this field excel and develop their career skills for positions both inside and outside academia...Resources at Addgene Read Our Posts on Networking Get Career Advice from Addgene Bloggers Learn How to Get Involved... career...

...Resources at Addgene Read Other Careers Posts Visit the Addgene Careers Page Learn about Management for...academia? Did I run screaming from the lab like the cartoon on the right?!? I would love to tell such a dramatic... a PhD and even more glad that I thought about careers outside of academia during my graduate training...you start thinking about preparing yourself for a career while still in graduate school, you’ll be well ... career development...

...demonstrate this quality.   I am often asked to give career seminars on Leadership Skills. After attempting...leadership different from good management? Aren't all career skills leaderhsip skills when done well? Do you...know it when you see it? “The Three Stages of Your Career: Doer, Manager, Leader” by Charlotte Beers, divides...these very different roles. At any stage in our careers we may be in a cycle of doing, managing or leading...spending a lot of time in doer mode even late in our careers.  Managers make sure the work gets done Managers...happy with a primary role of doer for your entire career. Scientists train as doers and often maintain an...

...two plasmids: pCas9: carries Cas9 and chloramphenicol resistance pCRISPR: carries a spacer targeting the... of interest and kanamycin resistance E. coli carrying the phage recombineering machinery are first electroporated...only 2-3 hours incubation. E. coli and T. citrea scarless editing plasmids Sheng Yang’s lab describes a ...recombineering with CRISPR to create a system for scarless, iterative genome engineering (Jiang et al., 2015...Gram-positive bacteria Jason Peters, Oren Rosenberg, and Carol Gross used two different genetic systems, targeting...10.1021/sb500351f Dong C, Fontana J, Patel A, Carothers JM, Zalatan JG (2018) Synthetic CRISPR-Cas gene...https://doi.org/10.1128/mbio.00019-20 Singh AK, Carette X, Potluri L-P, Sharp JD, Xu R, Prisic S, Husson...

... Courses HHMI Resources for Early Career Scientists Science Careers: Running Your Lab (pdf) Advice for... next professional adventure begins. While many career pathways exist for scientists and engineers, a ...to take things slowly! It is more important to carefully select first hires who are well-suited, productive...for starting your academic lab, download Science Careers' "Running Your Lab" booklet. What has been the...Institute (HHMI) offers some great resources for Early Career Scientists, including their "Making the Right Moves...

...Resources on Addgene.org Check out our careers page Watch our career videos ... school and beginning to research non-academic careers, I always felt that descriptions of transferable... this skillset. After sifting through dozens of career advice articles, I settled on defining problem ...to research and the lab, you may not be able to carry out the entire problem solving process on your own... and effectively will help you grow in whatever career path you choose.    Resources Additional resources...

...find those that do not carry a given dominant disease-causing gene, or that carry only one copy of a recessive...mutation in MYBPC3 that leads to hypertrophic cardiomyopathy. An adult male with a heterozygous 4 bp deletion... wild type allele (Figure 1). 72.4% of embryos carried two wild-type copies of MYBPC3 (versus 47.4% of...of untreated controls.) 27.6% of embryos carried indels at the mutation site, indicating that repair proceeded...many situations. In the case of hypertrophic cardiomyopathy and other genetic disorders, pre-implantation...