...already be seen with things like DNA synthesis, Golden Gate cloning, and CRISPR. The yeast synthetic genome ... He returned to academic research in 2005 to investigate synthetic biology at Boston University, working...yeast synthetic genome project. His team also investigates bacteria and extremophiles for biotechnology...Education Nonprofit BioBuilder Plasmids 101: Golden Gate Cloning Learn About Working at a SynBio Startup ...

...Addgene blog Plasmids 101: Golden Gate Cloning High Complexity Golden Gate Cloning with NEB’s DAD New MoClo...Drosophila. Similarly, the Multi-Kingdom (MK) Golden Gate system was designed to work in fungi, bacteria, ...s12896-017-0393-y. Engler, Carola, et al. “A Golden Gate Modular Cloning Toolbox for Plants.” ACS Synthetic...Chiasson, David, et al. “A Unified Multi-Kingdom Golden Gate Cloning Platform.” Scientific Reports, vol. 9, no...

...our growth over the last few years. Some of the navigation was buried on the page, and a lot of the content...clear to us that we needed to improve search and navigation, while also updating brand messaging, as well...this we’ve also made improvements to our global navigation and menus over the past year, making it much ...sharing, and fun Along with improving search and navigation, we also wanted to emphasize the values that ...

...overhang sets for high complexity Golden Gate Reactions Cloning ligation reaction calculators These links can... For general primer design Design primers for Goldengate cloning   Design primers for In-Fusion Cloning..., Gibson assembly, or In-Fusion cloning.  NEB Ligation Calculator: This tool works for any cloning method...

...voltage indicator: JEDI-2P Novel class of light-gated potassium channelrhodopsins Myc-tag antibody anti-c-Myc...10.1016/j.cell.2022.07.013.   Novel class of light-gated potassium channelrhodopsins by: Mike Lacy New work... John Spudich identifies a novel class of light-gated potassium channelrhodopsins, named Kalium channelrhodopsins... al. Kalium channelrhodopsins are natural light-gated potassium channels that mediate optogenetic inhibition...

...is directly conjugated to your reporter circumvents the issue. However, directly conjugated primary antibodies...antibodies. The good news: there are strategies for mitigating this issue, and we’ve highlighted a few below...endogenous IgGs by pre-treating the tissue with unconjugated Fab fragments that recognize the endogenous ...

...exclusively in E. coli? Gram negative bacteria in general? Both Gram negatives and Gram positives? Will you...has to do with the balance between positive and negative regulation and can be manipulated with mutations... strain Most E. coli strains can be used to propagate plasmids, but endA- E. coli are best for high ...

...study 1: Cell lines Many of the reagents used to investigate SARS-CoV-2 were developed for broad use and are.... Seventeen of these cell lines were used to investigate the susceptibility of cells from different organisms...choose the best experimental model for further investigation of this pathogen. The sharing and distribution...research institution, has been able to rapidly propagate this strain, making it available to scientists...

...including: Gibson Assembly®, BioBrick® Assembly, Golden Gate Assembly, and NEBuilder® HiFi DNA Assembly. Gibson...rational and simple shuffling of DNA regions. Golden Gate Assembly also allows the creation of standardized...polymerase) is the ability to remove, reset and ligate mismatched overhangs, yielding scarless DNA assembly...Resources on the Addgene Blog Learn More about Gateway Cloning Gibson Assembly and Other Long-Homology...

...color-changing flower, but to do that we’ve had to navigate the challenges involved in plant synthetic biology...unique metabolic processes of plants are being investigated on limited funding and time. While institutions...cloning kits available in the repository, MoClo and GreenGate, and we are excited to see how this exchange of...of potential, but there are several hazards to navigate when mapping out a new project. We’ve shared some...

...to cut out the inserts followed by a five-piece ligation with the four inserts and the master plasmid in...Addgene: Tell me about the history. How had investigators done this kind of work before? Musunuru: In ...Unfortunately, it is time consuming and difficult for investigators without a lot of expertise to create zinc finger...and puts it in the hands of pretty much all investigators. Our kit makes it extremely simple to design...

...Alistair McCormick’s lab recently developed the CyanoGate Kit for use of the MoClo system in cyanobacteria...PCC6803, and the fast growing strain Synchococcus elongatus UTEX2973, showing the potential for broad use.... Figure 3: Assembly of plasmids using the Cyanogate Kit and subsequent transformation into cyanobacteria...Image from Vasudevan et al., 2018. Find the CyanoGate kit at Addgene. Vasudevan et al., Plant Physiol...

....e. restriction cloning, Gibson Assembly, Golden Gate etc.), and try to find the simplest way to do it...Transformation Transform a few microliters of your ligation reaction. Here are the possible outcomes: No ...PCR, putting special attention in the areas of ligation between the different fragments If you don’t ...t get any positive clone, try transforming the ligation mix in different bacterial strains (stbl2, NEB...

...Library, deposited by Carrie Bhang, a research investigator in the In Vivo Pharmacology group at Novartis...the parent cell population using the library, propagated it, separated it into roughly identical replicate...ClonTracer Since starting her new role as an investigator in the In Vivo Pharmacology group at Novartis...Addgene community! Carrie Bhang is a research investigator in In Vivo Pharmacology at Novartis Oncology...

...timeline of the Golden Gate (GG) cloning approach. Individual modules and a sample ligation are depicted. (Bottom...assemble them into a destination vector via Golden Gate cloning, perform an exonuclease cleanup, transform...Cloning Addgene's CRISPR Guide Plasmids 101: Golden Gate Cloning...

...follow through on their promises, Wet-blanket negativist, who can bring down the morale of a group with...Some supervisors might try to intimidate you with negative comments (hostile-aggressives, complainers, know-it...know-it all experts, wet-blanket negativists, and micromanagers), while other supervisors might be nice...complete your work. Whether your supervisor is a negative person or just an inexperienced manager, you can...

... iodixanol gradient ultrafugation Overview: Iodixanol gradient ultracentrifugation uses a gradient of ...during AAV production is overlaid. Following ultracentrifugation, the AAV-containing 40% fraction is collected...day. Refer to the AAV Iodixanol Gradient Ultracentrifugation Protocol for more details. Pro tips Watch...

...days Purification Ultracentrifugation (AAV); centrifugation (lenti) Centrifugation Titration time ~...is typically purified using methods like ultracentrifugation, while lentivirus supernatants are centrifuged...

...detect mEPSPs, it also enabled imaging of their propagation along dendrites for the first time (Figure 3)...without the Kv motif. Altogether, if you want a negative-going GEVI that can be used in different illumination...include large-scale pharmacological profiling or interrogation of context-dependent signaling in primary cells..., J. C., & Garcia-Marcos, M. (2024). Direct interrogation of context-dependent GPCR activity with a universal...

...interest. Select for cells of interest (positive or negative selection). Next-generation sequencing (NGS) to...high degree. This is particularly helpful when investigating tissue-specific diseases, as you can target ...protein altogether. Platforming AAV Interest in interrogating and mapping the genetic landscape of tissue-...used to map tumor suppressors in mouse liver, interrogate immune cells to improve immunotherapy in glioblastoma...