We narrowed to 662 results for: Clu

...chemically distinct subcellular environments, including the organelles that compose the secretory pathway... significant post-translational modifications including glycosylation, disulfide bond formation, and proteolytic...real consequences of misfolded dark FP fusions including erroneous quantitation of fusion protein levels...imaging in a variety of cellular compartments including the secretory pathway (Figure 2), the inner membrane... Anatomy and Structural Biology. His interests include the quality control of secretory proteins in the... PMC2172584. 11. Jokitalo, Eija, et al. "Golgi clusters and vesicles mediate mitotic inheritance independently...

...experiment lacking the proper controls are invariably inconclusive and unreliable. Proper controls provide the ...human cells. From this result one could simply conclude that the shRNA didn’t work, as the expression ...control plasmids Part of planning your experiment includes determining what factors need to be controlled...microscopy images resulting from our experiment that includes the aforementioned internal control and is consistent...to note that optimal experimental conditions, including how much plasmid DNA to use for any individual...plasmid-based experiment, untreated cells should be included as these provide the baseline/standard against...

...Project conducted a large scale sequencing and concluded that nearly all disease-linked human genes have...as limbs, the spinal cord, and internal organs including the heart, brain, and lungs (McCusker and Gardiner...constructed several vectors for use in the axolotl including plasmids for use in CRISPR and Tol2 transposase...Some of the most popular tools for studying frogs include egg transplants and microinjections. Injections...fluorescent fusions in a multitude of organisms including Xenopus. Overall, the ability for scientists to...function. Some key genetics research with Arabidopsis include understanding of allelic variation between different...

...to record keeping. Top priorities in most cases include IP protection, publications and funding. If any...which focused on the adoption of ELNs in labs and included hundreds of researchers. The full story is available...was a significant barrier to ELN adoption. This includes financial outlay, staff hours, troubleshooting...emails. Important collaborative functionalities include comments, notifications, sharing files, assigning... clients, you might need share data with them including reports and protocols. To facilitate this, some...meetings with the support teams behind them.  Conclusion Researchers spend 40% of their time on tedious...

...traditional restriction enzyme cloning we once relied exclusively on. In this blog post, I will go over some advantages...N-terminus, but you don’t have the tag already included in the vector you want to use. If you inserted...primers to amplify the two fragments while also including regions of homology to the vector or neighboring...no need to screen a large amount of colonies. Excluding the time it takes to get primers, you can have...sequences between fragments. Some of these products include: GeneArt® Seamless Cloning and Assembly (Thermo...enables one-step targeted cloning of large gene clusters. Nat Commun. 2015; Sep 1;6:8101. PubMed PMID: ...

...that there are many companies and organizations (including Addgene), that need people who are data oriented...academic environment. Some great tips from this panel included:  Write Often Whether you take on opportunities...publishing process. The diversity panel - Equity and inclusion The final and possibly the most important panel...show that people from all backgrounds are truly included as members of their research communities and are... be asked going forward is: How do we measure inclusion and equity? This is a difficult question but not...example of how underrepresented individuals are discluded/not treated equitably even when hired; well-meaning...

...systems and creating an all-synthetic vector that included only the elements needed for function. The lab...channelrhodopsins for use as optogenetics tools. These tools include a pair of channelrhodopsins, Chronos and Chrimson...CsChR, and CoChR have been deposited to Addgene, including lentiviral and AAV expression vectors for certain... for the Rinehart phosphoprotein system, which includes detailed protocols provided by the Rinehart lab...years of distributing his Designer Receptors Exclusively Activated by Designer Drugs (DREADD) plasmids...repressor plasmids from Scot Wolfe's lab. These include Tet-inducible CRISPR activators and repressor plasmids... contain a variety of N- and C-terminal tags (including His, S-tag, OneStrep, CBP, Trx, GST, Halo, MBP...

...create knockout alleles in a variety of species, including the standard laboratory mouse. It’s also made ...bacteria. But CRISPRing typical mouse models, including creating Cre-dependent conditional alleles, has...gRNAs. A variety of other undesired outcomes, including one loxP site insertion and one NHEJ-repaired ...problem. Briefly, you design your donor template to include a T7 promoter upstream of the left homology arm...your ssDNA. Note that the T7 promoter will not be included in the final ssDNA template. Figure 2: ...single-stranded DNA production services or kits, including the IDT Megamer service tested by Quadros et al...

...plasmids are divided into two sets, each of which includes plasmids with different varieties of tags, both...quickly and easily. These tagged plasmids also include cleavage sites should one need to remove the tag...Bacillus subtilis and Pichia pastoris pCri also includes plasmids designed for protein expression in systems... coli. Plasmids designed for these systems may include a signal peptide (SP) that routes the protein through...can improve disulfide bond formation. pCri also includes a special feature for difficult-to-purify membrane...

...dataset currently includes 1049 community-validated reagents from 20 contributors, including 61 fluorescent... That’s why scientists from around the world, including Andrea Radtke, PhD, and Ziv Yaniv, PhD, have created...entire dataset can be downloaded from Zenodo, including the reagent resources .csv file for easy filtering... And the dataset is rich. Experimental data includes reagent type, target species, target name, host...

...users were included in the dataset; 2) users not part of a scientific Twitter list were excluded from the...follow each other, or are friends, are included in this cluster analysis. The closer two dots are to each...(blue) and social (orange)  scientists largely cluster by discipline, suggesting that scientists tend ...

...reagents for many molecular biology protocols, including vector-insert joining for recombinant plasmid ...efficiently ligate many undesirable structures, including substrates containing gaps or DNA base pair mismatches...activity, and for others, there is a need for the exclusive ligation of fully base-paired nicks with no gaps...specificity and quantitative accuracy and even include profiling single nucleotide polymorphisms (SNPs...substrate (23, 24). Thermostable DNA ligases, including Taq DNA ligase, are naturally able to discriminate...DNA ligase commercially available (see below). Conclusion It is important to note that thermostable, high...

...its associated partners. Common pull-down assays include immunoprecipitation mass spectrometry (IP/MS) and...for each cell type you work with (optimization includes selection of transfection buffer and electroporation...assays using a tag-specific antibody.Note: Always include a positive and a negative control for the tagging...measure of ICC assay specificity. We generally include Sox2-V5 tagging as a positive control in our knock-in...experiments depends on many factors. Some of these include: Most importantly, the crRNA should cut near...Medicine (CRM), Edinburgh. His research interests include optimizing CRISPR/Cas9 genome-editing methods and...

...the tool to display excitation spectra, which I excluded here for better visibility. Source: https://www.bdbiosciences.com... in Table 2 is a standard and should always be included—the live/dead stain. This stain is in fact not... cytometer's software. It also means we should exclude APC-Cy7 from the rest of our panel. Besides some...allow us to detect the eGFP signal. We will now exclude FITC from the rest of our panel. Lastly, we want...www.bdbiosciences.com/en-us/resources/bd-spectrum-viewer.   Conclusion You made it! In this post, you learned about ...Sharpe at Harvard Medical School. His projects include investigating negative regulators of T cells and...

...to engage the local community (including local garden and compost clubs) to participate in this study....science projects, we offer classes, workshops, book clubs, seminars, and social events. Building a community...reactivate it. We started out by holding a few journal club events and soon realized that it was easy to find...people who were interested. For example, our journal club discussion on Kevin Esvelt’s CRISPR/Cas9 gene drive...

...things you need to include in your manuscript. What information do I need to include in my paper? More ...and your results more valuable. Some journals, including F1000Research, also ask for the raw data behind...good quality and whether they agree with the conclusions of the article. These initiatives are another... to your submission, briefly summarizing the conclusions of your paper. This will help the editor handling...

... that come with working with prokaryotes. This includes but is not limited to: rapid growth, ease of replica...bacterial cell. Similarly, the specific ORI elements included within a yeast vector determine how the plasmid...controlled if specific regulatable elements are included (reviewed here). Plasmids for use in S. pombe...Scientists can exploit these host mutations by including a copy of a functional gene which complements ...extensive list of yeast auxotrophic markers and includes the associated references.  Yeast selection...

...this specific conference, and I was able to include and exclude different modules for this workshop. Empowering...example, Addgene’s contributions by Tyler Ford included reagent sharing and were presented at the ASPB...learned: Since early 2018 the team has grown to include experts who could help provide new materials and...assembled the right people at the right time. Be inclusive and seek solutions by working with communities...

...Constantini, 2012), whereas it was previously concluded to be monomeric based on gel filtration (Chudakov...of plasmids that can be used in the OSER assay including CytERM-mKOkappa, CytERM-mTurquoise2, and CytERM-dTomato...for proper ER structure. Based on these data, we conclude that mKOk has dimerization tendency and is not... acts as a monomer in the cell type tested. To conclude, we recommend that you try a couple of different...established true monomeric fluorescent proteins. Concluding remarks Many fluorescent proteins are available...

...reference pages and curated databases for scientists including Wormbook, Wormatlas and Wormbase. And scientists...workshops not only highlighted new tools but also included time for questions and a group discussion on the... to view and explore the worm. These resources include the open worm browser, which allows you to view...elegans locomotion. To see the other resources included in OpenWorm, check out the OpenWorm website. microPublication...microPublications is curated and added to other databases including wormbase and flybase. microPublication is run ...

...experimental manipulation. Common inbred strains include C57BL/6, Balb/c, and 129. Conversely, outbred strains...genetic engineering. Examples of outbred strains include Swiss Webster and Harlan (Hsd) National Institutes... such as an antibiotic resistance gene is also included to select for successfully modified zygotes or...Targeting vectors used to create knockout mice include a reporter gene, which replaces the targeted gene...floxed sites in the cell can occur. Similar systems include tetracycline or doxycycline mediated activation...

...The pegRNA consists of a CRISPR gRNA extended to include a primer binding site (PBS) complementary to the...genome and a reverse transcriptase (RT) template including the desired edit. Figure 1: Prime Editing...BioRender.com. RNA protectors epegRNAs: epegRNAs include structured motifs at their 3′ end to protect them...binding site (Nelson et al., 2022). PE7: This editor includes the fused RNA-binding protein La to protect the.... That’s all our tips for today! Each tip includes a citation to the paper it was drawn from. If ...

...These tables are not inclusive of all methods, nor are these methods limited exclusively to in vitro cell ...are required for any application. If you want to include a homology directed repair (HDR) template to create... table below summarizes the various components included with each expression system as well as suitable...components into mammalian cell lines is quite broad and includes several different methods, so we've summarized...

...Gate Modular Cloning (MoClo) Kit. What does it include? Patron: Sainsbury has a mandate to share and educate...things for day-to-day life in a plant lab, so it includes everyone’s favorite promoters, plus a few new ...resources. We wanted to expand the toolkit. We have included a wider range of terminators than are commonly...a modern toolkit for those working in plants, including many things people already use. It makes the switch...Addgene recently to explore their use in plants, including FRET tools, double dimerization GFPs, and circular...

...will discuss later.       Standard curves must be included every time the assay is run, ideally with freshly...curves vary depending on a number of factors, including the user, equipment, assay incubation time, etc...curves must be validated for each run. To validate, include a positive control of known concentration in the...Dynamic range depends on a number of factors, including the dilution series of the standard and the minimum...account the range of concentrations possible.  Include both a standard curve and a positive control in...

...screen for molecular interactions in the cell, including protein-protein, protein-DNA and protein-RNA interactions...reporter gene (3, 4), there are now many variations, including many that use auxotrophic rescue genes as reporters...likelihood of indiscriminate reporter gene activation. Including a prey-only control also provides a baseline level...reporter signals depend on a number of factors, including protein expression and correct folding, post-translational...-hybrid systems available in model organisms, including bacteria, alternative fungi (C. albicans, pC2HB...

...interactions. For difficult to release proteins, including those in the nucleus, more stringent buffers may... may be needed. Since many factors, including salt concentration, ionic strength and pH, affect binding...may degrade the target protein. To prevent this, include protease and phosphatase inhibitors in the lysis...Controls As with any experiment, it is critical to include proper positive and negative controls for an IP...sample that does not. Common negative controls include knockout cell lines or tissues that do not express...

...spacers of the crRNA can be customized as well, including concatenations into crRNA arrays targeting multiple...by sacB counterselection.  The MUCICAT toolkit includes versions of plasmids with different induction ...terminators can serve to interrupt genes (or gene clusters) in metabolic engineering. So far, we’ve used ...concentration. Image from the Yang lab.   In conclusion, the MUCICAT system has the following advantages...engineering with improved cargo dedicated to gene (cluster) interruption.     Plasmids for each of these...

... the 1930’s in a range of biological studies, including investigations into environmental pollutants and...organism with a clearly established body plan including head, eyes, and tail. Compared to the opaque mouse...process. Additional zebrafish applications and conclusions Zebrafish are also popular outside of developmental...not ideal for every application. Disadvantages include their duplicated (i.e., redundant) genome, their... these caveats, several features of zebrafish—including the available genetic resources and transparency...

...that went beyond obtaining “omic” data and had to include clear plans to connect Genotype to Phenotype thus... is to ensure that, in general, these labs all include materials sharing in their dissemination plans ... diversity in the social bonds that they form including genetic mechanisms that control who a female wants...of basic evolution of fungi and eukaryotes and includes the fungus that is killing frogs all over the ...that most cephalopods die after they lay their 1 clutch of eggs? Propagation is thus quite difficult. CRISPR...

...non-functional hedgehog, the denticles actually clump together forming one patch on the surface, giving...contain components of the SB transposon system including those from the original publication, Ivics et ... been studied in many contexts within the fly including oogenesis and cytokinesis. Why name this protein...activate bagpipe which is localized to segmental clusters of the mesoderm. This subdivision of the mesoderm...that contains bagpipe from the Deplancke lab. Conclusion I hope you enjoyed this tour of Addgene’s new...

...multiplexed elements. The team designed sgRNAs that include an internal barcode and separated multiple sgRNAs...was first described in Anzalone et al., 2019, including modifications to the Cas9 or reverse transcriptase...coding sequences for a range of applications, including CRISPR activation by dCas9-VPR, prime editing,...REVeRT in various contexts in vitro and in vivo, including in human retinal organoids and live mice. Importantly..., Parkinson’s, ALS, and Huntington’s Disease, including plasmids, antibodies, viral vector preps, CRISPR...

...depend on target and sample type. Example steps include: permeabilization, antigen retrieval, and sectioning...available, and your experimental goals. Example steps include: counterstaining, mounting sample on slides, and...couple of steps are preliminary steps that are not exclusive to IF, but they can have a significant impact ...you optimize your protocol, always remember to include controls to show that your antibodies are binding...antibodies validate them in a variety of applications, including IF (or IHC or ICC), and will share some of their...

... blue light CRY2-CIB1 heterodimerizes or forms clusters under blue light Check out our Optogenetics Guide... One great way to do this is to use CRY2-CIB1 clustering. Fuse either CRY2 or CIB1 to your protein of ...Rab protein vesicle trafficking via CRY2-CIB1 clustering. Image adapted from Aljiboury et al., 2023, under...earlier work on Rab11, the team used optogenetic clustering of Rab8, Rab11, and Rab35 to understand their...attractive strategy for engineering other enzymes. Conclusion We hope this post has illuminated some of the...

...editing to change the genetic code of plants, including the development of crops with longer shelf life...to engineer the genome of many plant species, including commonly used model organisms like Arabidopsis... Medicago truncatula and several crop species including potato, corn, tomato, wheat, mushroom, and rice... used RNA pol II promoters for Cas9 expression include the ubiquitously expressing cauliflower mosaic ...toolkits for creating plant expression plasmids, including plant CRISPR plasmids from the Yiping Qi lab and...

...Symptoms of AIDS include rapid weight loss and an increased risk of infection, including both common infections...at increased risk for other chronic diseases, including dementia, gut disorders, nerve damage, and cardiac...could be used to combat this problem, just as ART includes multiple drugs to lower the odds of developing...near the Cas9 cut site, leading Wang et al. to conclude that some Cas9-derived indels may not ablate viral...

...workplace bullying? Threats to professional status – including belittling opinions, public professional humiliation...competence procedures Threats to personal standing – including undermining personal integrity, destructive innuendo..., intimidation, sexual harassment Isolation – including preventing access to opportunities, physical or... withholding necessary information Overwork – including undue pressure, impossible deadlines, unnecessary...unnecessary disruptions Destabilization – including failure to acknowledge good work, allocation of meaningless...

...technology can be used for numerous applications, including transgenesis, gene-trap screens, and gene editing...manipulations in animal models or cells lines, including stem cells, without leaving behind any accessory...piggyBac™ transposon containing a selection marker is included in a standard homology directed repair template... used for a variety of research applications, including making transgenic cell lines and genetically-modified... using a variety of gene editing technologies including Zinc Fingers, TALENs, and CRISPR-Cas9 nucleases...

...high as 1-3%. In light of this finding, we now include guidance on the plasmid page of DIO/FLEx AAV vectors...you intend to use performs in your system. This includes a thorough testing of both the serotype and promoter... is reached you may obtain a false negative or conclude the viral vector is not working.  When using a...off-target expression that can confound your results.  Conclusion Using AAV vectors in your experiments requires...system. It is very rare for a methods section to include all of the variables that need to be accounted ...

...Wilson and Doudna, 2013). RNAi consists of two non-exclusive gene silencing pathways: transcriptional and post-transcriptional...afflicted by pervasive off-target effects. This conclusion was based upon numerous phenotype discrepancies...importance of using orthogonal methods to confidently conclude the function of a gene of interest. Tools for ...discussed below). Examples of these constructs include pGAPi, which allows easy creation of long hpRNA...experiment are now available on Addgene. These include the construct that allows for easy insertion of...