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...researchers can measure this conformational change to investigate the role of specific integrins in cellular processes... αL, αM and α5. IPI’s anti-integrin rAbs are unconjugated and recommended for use in flow cytometry. Based...enable cells to spread, resulting in cells that elongated and morphed asymmetrically (Jo et al., 2022). ...subunit. Using these affinity reagents, the investigators inhibited other RGD-binding integrins, individually...for cell adhesion. The results surprised the investigators, challenging the long-held belief that αVβ3 ...

...control We will discuss three types of tools: light-gated ion channels, optogenetic G-protein coupled receptors...receptors (OptoXRs), and photoswitches. Light-gated ion channels directly open and close when exposed to... inhibit a specific receptor or pathway.  Light-gated ion channels Light activated ion channels (rhodopsins...sustained period of time (Gong et al., 2020). Light-gated ion channels are useful when you want to activate...E. (2003). Channelrhodopsin-2, a directly light-gated cation-selective membrane channel. Proceedings of...

...of citable, linkable figures. Cause #2: Hidden “negative” data We all want to be able to tell a good story...however, made it difficult to publish so-called “negative” results that conflict with our original hypotheses...problem would be for traditional journals to publish negative data, but this is a big ask and, from the journal...:31 - 49:45), there are many ways to publicise negative data - we don’t have to rely on traditional journals... whether the ultimate results were positive or negative. I can feel you rolling your eyes, but even if...

...constitutively-active or a dominant-negative mutant. Reducing the copy number can also mitigate the deleterious effects...expression of certain genes, such as those that have negative impacts on fitness. One consideration when making...enzyme [4]. By using a donor plasmid containing a negative selection marker such as thymidine kinase, a researcher...efficient as Cre recombinase. However, the use of negative selection largely overcomes this obstacle. Furthermore...

...different likelihoods of returning false positives/negatives, but they all generally yield a similar result...s test – Most conservative test. Higher false negatives, lower false positives. Newman-Keuls method –... – More powerful. Moderate false negatives, moderate false positives. Duncan’s multiple range test – Most... Most powerful. Low false negatives, high false positives. Control vs. all samples Oftentimes there exists...risk of higher error, both false positives and negatives (or, as the mathematicians say, Type I and Type...

... in popular fiction has been largely negative. But these negative depictions serve as cautionary tales...the information necessary for regulating and propagating life, there was no turning back. While the world...wreaking havoc in a theme park.  Released in 1997, GATTACA is an American SciFi film starring Ethan Hawke,...between Cas9 and reverse transcriptase. Using an elongated template + guide RNA called pegRNA (prime editing...

...Hendel et al. 2015). Low detection rates mean the investigator can be relatively certain that their samples...off-target mutations are essential data points for investigators looking to definitively link genotype and phenotype... chromatin configuration, produces many false negatives Combining sequencing techniques can ensure ...genome-wide unbiased methods. However, most investigators use single cell clones for in vitro CRISPR experiments...gene editing achieved by TALENs, CRISPR/Cas9, and megaTAL nucleases." Molecular Therapy (2015). PubMed PMID...

...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...

... post was written by Professor Mark Howarth, investigator at the University of Cambridge. His research...expression C234-E25: attB2r-SNAP-attB3 SNAP-tag Gateway entry clone to add C-terminal SNAP-tag Promega...template VirD2 HUH-tag For creating DNA-protein conjugates rHUH HUH-tag (RNA) Covalent protein tag for...cellular computation, making possible multiple logic-gated steps. Beyond the lab, these systems can also assemble...

...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...