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...ends, some 50% of these groups continue to meet regularly, providing ongoing support, advice and accountability...and mentees. Because the participants are at a similar development stage, they have a lot to learn from...Sometimes it is helpful to find peers that share a similar direction or problem. For example, a group of people...most hits. Commit to a meeting schedule and meet regularly no matter what, even if one person can’t make ... work at the same place, a 1-2 hour lunch on a regular schedule might work. If not, perhaps a 3 hour breakfast... handy list of suggestions.  Get started. Have regular meetings. Experiment with the process. Have each...

... we will focus on a unique gene that makes the popular GatewayTM method possible: ccdB. But what is ccdB...start with a brief history of the gene and how molecular biologists have harnessed it to evolve cloning...clones.   ...Becomes a powerful cloning tool Molecular biologists first saw the potential of this system...and are considered F-); however, there are a few popular lab strains, such as NEB Stable, JM109, XL1 Blue...experiments. Please check out Addgene’s Choosing a Molecular Cloning Technique page and the Addgene Blog for...Cloning Posts Resources on Addgene.org Choose a Molecular Cloning Technique Find Plasmid Cloning Protocols...

...described above can be used for similar applications and have similar advantages, though with some caveats...chain. Created with biorender.com     The modularity of antibodies means that it is relatively easy..., two full antibodies bound together may be too large. Using a complex that is one full antibody bound...Alternatively, Fab fragments can be used as a block in similar scenarios. By applying unconjugated Fab fragments...and have lower binding affinity than some of the larger fragments.  Think of antibody fragments as new ...

...: IL-36 biology in generalized pustular psoriasis Generalized pustular psoriasis is a chronic autoinflammatory...in this educational video. “SLC6A8 belongs to a large family of solute channels that are responsible for...outside of the cell and translate those signals into cellular responses. For example, GPCRs and their downstream...different receptors upon stimulation. “To study cellular transduction events, we will utilize BRET-, FRET...intensity-based signaling biosensors to monitor intracellular signaling,” says Hébert. Previous research has...

...reagents.  Affinity Reagents  Affinity reagents are molecular tools that have the ability to specifically recognize...affinity reagents is their size. Antibodies are large enough that their bulk can be a barrier at times...difficult to source, may not be readily available for a large number of targets, and can require specific conjugates...   Types of Affinity Reagents  (Listed in no particular order.) Fab fragments are created through enzymatic...variables (scFvs) are ~27kD engineered fusion proteins similar to a Fab fragment but created through engineering...eukaryotic cells, allowing for them to be used as intracellular inhibitors.  Aptamers are not proteins at all...

...time intensive to process a large number of microbiome samples. To enable large-scale, longitudinal studies...Gram-negative cells.  The Wang lab further developed a modular suite of plasmids (pGT) with various replicative... honey bee microbiome. The toolkit consists of modular parts that are assembled with Golden Gate Assembly...other microbiome systems. Find the toolkit for modular assembly of broad host range plasmids Counting ...bee gut microbiome bacteria with a toolkit for modular assembly of broad-host-range plasmids." ACS synthetic...

...potently inhibit type II CRISPR systems including the popular S. pyogenes Cas9 (Rauch et al., 2017, Hynes et ... on a single plasmid (Grefen and Blatt, 2012). Similar to traditional Gateway™ cloning, the GOI first ... (FRET, Heckler et al., 2015) or ratiometric Bimolecular Fluorescence Complementation (rBiFC, Grefen and...offers a way to spatially and temporally control cellular processes in vivo. Many photoactive proteins exist...photo-reversible protein-protein interactions can control subcellular localization processes in vivo. They deposited...the mitochondria, because cPYP binds to BoPD. A similar set of newly identified light- and dark- binder...

...can be particularly useful in academic labs when creating shared experimental protocols. Clarify: Asking...setting, where student and postdoc turnover happens regularly and it behooves everyone in the lab to be using...Asking for clarification during a meeting can help get everyone on the same page. Others in the meeting might...communication has helped Addgene move from completing 2-3 large projects a year, on average, to completing 8-10 ...understanding you can be incredibly frustrating. Clarifying the words —> decreasing frustration —> increasing...

... to interact by stimulation with blue light.  Similarly, light can also be used to control homo-oligomerization...distribution of clusters without accumulation in subcellular compartments such as nuclear speckles. Furthermore...OptoSTIM1), caused two-fold faster changes in intracellular Ca2+ levels than the original version containing...CRY2clust should prove a useful tool to further probe cellular function. The CRY2clust Plasmid can be found here... commonly observed with direct stimulation are largely absent in the soCoChR-expressing system. Taken ...interactions. We’re excited to see how you adapt similar FLINC biosensors to work in your lab. Find the ...

...encode proteins. They play important roles in many cellular and regulatory processes, but much of their functionality...removing the full ncRNA locus (Ho et al. 2012). For large-scale screening, sgRNA libraries are useful for ... same time (Liu et al. 2018). However, removing large lncRNA fragments with multiple gRNAs poses the risk...Zibitt et al. 2021). Mutating splice sites is particularly effective as it can result in the retention ...Mutations of splice sites. (C) Removal of an exon or a large genomic fragment. (D) Knock-in: insertion of a synthetic...used to target lncRNAs with high specificity, particularly in applications requiring smaller delivery systems...