... passion, and patience are required to build and lead a productive and successful research lab. Once goals...champagne, that's a given.” Furthermore, anyone leaving the lab receives send-off via a gift, card, and... the afternoon during the child-rearing years, learning “lots of interesting things from children, their...their classmates, and parents”. He also cooks and cleans with his family, “sometimes listening to audiobooks...

...functionality can be found in the following video: Please remember that the accuracy and reliability of our... plasmid is missing from our collections pages, please shoot us an e-mail at [email protected]. We’d love...visibility in a few other ways. Every quarter we release a Newsletter where we spotlight some of the plasmids... in the repository. And last, but certainly not least, we often feature new plasmid technologies on our...

... also have the option to “Leave us a message.” When you click on the "Leave us a message" button shown...lab’s materials and requesting them from your colleagues, to intellectual property rights and handling...the Addgene Blog Read other Inside Addgene Posts Learn about New Features on Our Plasmid Pages Resources...

...insights discoverable. Watch this quick video to learn how ReFigure works and continue reading for more... and again as they don’t save the insights they glean from a reading session. To remember what happened...reach the ReFigure team at [email protected] Please support ReFigure by creating ReFigures and answering...video to share your science at We Share Science Learn more about Data Sharing Resources at Addgene.org...

... you want. But how do you know if a masters will lead to a career you will like? Here’s two things to ...setting up an informational interview with them to learn how their master's degree prepared them for their... help with manuscript and grant preparation, and lead your own research project.  3. Do you prefer being...eBook.   Resources Resources on the Addgene blog Learn about career planning resources What Do I Do Now...

... year. The society facilitates opportunities to learn about Synthetic Biology and exciting new technologies...network of fellow scientists that you can grow with, learn from, and mentor while pursuing your research interests... which would benefit from some Addgene support? Please get in touch, as we aim to support a couple of ...Networking Get Career Advice from Addgene Bloggers Learn How to Get Involved in a Profession Organization...

...more years with a small number of formal advisors learning how to be a scientist. It is shocking how little...important in both cases. As a PhD student you need to learn how to be a scientist. Your PI should be the kind...science career. Will you be given opportunities to learn and practice successful grant writing, lab budgeting...Harvard Medical School researcher Connie Cepko to learn about her mentoring style. How can I find out if...

...genome. Cas genes, which code for polymerases, nucleases, and helicases, were also disrupted to elucidate...CRISPR loci to generate RNA, which guide a Cas nuclease to the specified exogenous genetic material next...within a host cell. A few virus particles have learned to circumvent the CRISPR/Cas defense by generating...mutation in the PAM sequence, preventing the Cas nucleases from re-identifying it (3). Surprisingly, few ...

...tertiary structure when transcribed which aids in nuclear export of mRNA and helps increase the expression...The polyadenylation or polyA signal aids in the nuclear export of RNA and RNA translation, and promotes....2019.276 Additional resources on the Addgene blog Learn more about basic viral vector elements Read about...about four considerations for the perfect AAV tool Learn about synthetic promoter AAVs for cell-type expression...

...indicated that a reasonable rule is to require at least 96% sequence identity when detecting a reference...commercial plasmids were inconsistent and sometimes clearly wrong. In an effort to be rigorous, we dug into... harness common feature detection in a new way, please contact us at snapgene.com. Many thanks to our...Blog Read about our SnapGene powered plasmid maps Learn how we use SnapGene during the plasmid quality control...

...few quirks specific to the PCR cloning process - please check out that post if you need a more detailed...PCR primers for molecular cloning consist of: Leader Sequence: Extra base pairs on the 5' end of the... existing software (a quick internet search will lead you to here and many others). If we put the sequence...product. Additional Resources on the Addgene Blog Learn About Other Cloning Techniques Catch Up on All Things...

...transferred between cells. At the time, these extranuclear agents of heredity were thought of as everything...first plasmid “cloning” experiment. Dr. Cohen and colleagues treated a tetracycline resistant plasmid, pSC101...topology, and cloning and gene expression. Just released: Addgene's Plasmids 101 (2nd Ed.) eBook And now...and more pictures and pro tips.  Happy reading! Learn more about the history of plasmids and cloning: ...

...transduction.) Compared to CCP transduction, iTOP is at least four times more efficient in primary fibroblasts... gRNA, which subsequently enter the nucleus and cleave the specified target. Figure adapted from D’Astolfo...expression is by nature transient, which may lower nuclease toxicity or the potential for off-target effects... Biol. 2000 Jul;10(7):290-5. PubMed. Is VP22 nuclear homing an artifact? Lundberg M, Johansson M. Nat...

...viral vectors. Here are some of the AAV we have released in the last few months. You can also browse our...coming soon! These vectors listed below should be released at Addgene in the next 6 months, pending quality... particular catalog item is available for order please email [email protected] with the vector name and...viral vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...

...accessible to everyone but these projects could also lead to novel yeast creations that may not be pursued... the different samples. The goal is to not only learn something about the microbial diversity in different... to find a community lab in a city near you. To learn more about what we’re up to or how you can help ...Disease & Open Science: Interview with Darrell Kotton Learn How Plasmid Sharing Speeds Science Resources at...

...of happen with Cas13. CRISPR-Csm degrades both nuclear and cytoplasmic mRNA with 90-99% knockdown efficiency...CRISPR-Csm presents an attractive alternative for nuclear-RNA targeting, where siRNAs are ineffective and...expression changes of non-target genes and RNA-seq of a nuclear (MALAT1) and cytoplasmic (CKB1) RNA target. Adapted...antibody per target just isn’t enough and we recently released Anti-GFP [N86/8R] as another option for targeting...

...interesting at an event. Or, perhaps you have a colleague who seems fun and helpful who you’d like to get...connections. I mention this because I want it to be clear that it is completely impossible for me or anyone...contacts over the years in many different ways. This leads me to my last tip: It’s usually not worth connecting...networking Attending a conference soon? Get tips here Learn about informational interviews ...

...embryo—even if it’s not necessarily via CRISPR/Cas9. This leaves us with the second critique. Absent of a broad ...intended parents is “rehoused” in a separate, enucleated zygote with healthy mitochondria, with the intent...Nuffield Council’s report is that when it became clear that this assistive reproductive technique was feasible...featuring a clinical pregnancy, was rejected by at least one academic journal for exactly the critiques leveled...

... work to enable future discoveries. Read on to learn more about Xin and Fatima's work and keep checking... work at the 2017 Winter qBio Meeting. You can learn more about Xin’s work with photoswitchable components...enzyme breaks down the HMO of interest, the lactose released activates expression of GFP from the lac operon...Biology! Additional Resources on the Addgene Blog Learn More about Photosactivatable Fluorescent Proteins...

...it unuseable. With the large size of the SpCas9 nuclease (~4.2 kb) this is a particularly prominent problem... perform an optimization-scale infection and at least one screening-scale experiment achieving the gRNA...the publications associated with these libraries. Please note, the pooled libraries we distribute in lentivirus...or are thinking about using our viral services. Please send any questions or comments to help@addgene....

...night of the iPSC symposium, a panel of thought leaders in the field--including Shinya Yamanaka, George... of ethics can be daunting because they have no clear cut answers, I applaud the stem cell community for...Disease; and Kevin Eggan presented work that is leading to promising clinical trials for ALS.   I have ...that!  Additional Resources on the Addgene Blog Learn about Efforts to Fight Rare disease with iPSCs Read...

...stakeholders to learn more about their needs and any problems that they are trying to solve. After learning more ...Technologies, I work with our customers in the US to learn where the gaps are in their sequencing workflows...problem-solver. A lot of the time, the problem to solve is clear, but the solution is more complicated. You’ll need...

...experiments. As a common example, a selection will leave you with the colonies that contain your plasmid ...negative controls for your PCR.  Find the details and learn more about colony PCR in this blog post.   ...can confirm the sequence of the entire plasmid. Please note that NGS verification is more time-intensive...biology protocols and tips Resources on Addgene.org Learn more about Sanger sequencing Read the Molecular ...

...are some of the AAV and lentiviral preps we have released in the last few months: Lentiviral preps for ...coming soon! These vectors listed below should be released at Addgene in the next 6 months, pending quality... particular catalog item is available for order please email [email protected] with the vector name and...viral vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...

...fluorescent reporter assays to quantify targeted Cas9 cleavage, and repair through homology directed repair (...while cells containing the reporter gene that was cleaved by Cas9 and repaired via HDR using the supplied... seamlessly remove the integrated GFP reporter leaving you with just your desired genomic edit.  Find ...using GEmCherry2. In this experiment they saw a clear difference in mCherry expression using FACs sorting...

...collection in your experiments and published your data, please fill out this form and we’ll add your citation ...combinations of WT, codon optimized, untagged, cleavage sites (thrombin and TEV), and C-terminal tags ...downstream experiments and offer flexibility in tag, cleavage, codon usage, and expression system. Mammalian...Addgene blog Read all of our COVID-19 blog posts Learn about other COVID-19 plasmids available at Addgene...

...Addgene blog Learn about other CRISPR methods for bacterial genome engineering   Learn more about lambda... novel genetic variants in bacteria. Read on to learn about the key components of EvolvR and its potential...extends it from the 3’ end, while its native endonuclease activity degrades the displaced strand.   ...

...ever like to write about a recent plasmid deposit please sign up here. Here's what you'll find in this...a continuation of that work, they have found at least two variants that display increased neuronal expression...microbial SSAPs (dCas9-SSAP) which insert genes in a cleavage free manner. The dCas9-SSAP system reports an ... less risky.   Fig. 3: Development of a cleavage-free dCas9-based gene editor using microbial SSAPs...

...ever like to write about a recent plasmid deposit please sign up here.  Here's what you'll find in this...of anti-FLAG-FB-GFP binding to FLAG-tag H2B for nuclear localization. (c) Applications of anti-FLAG-FB-...anti-FLAG-FB-GFP labeling various targets with nuclear (top, 4x-FLAG-mCherry-H2B), cytoskeletal (middle, 4xFLAG-mCherry-β-actin... editing, while gene editing can be done via a nuclease-active CasMINI. This small yet powerful system...

...option, in aliquots no smaller than 10uL. You can leave them at room temperature when you’re working with...illustrious lab career, you will likely accidentally leave your antibody out on the bench overnight. Do not...this post, the conjugates, or signaling molecules! Please keep in mind that this is just a brief introduction...data, you’ll want to pick fluorophores with the least spectral overlap between them whenever possible....