...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...

...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...

...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...

...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...

...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...

...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...

...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 ...

...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...

...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...

...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...

...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...

...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...