...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...

...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...

...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...

...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...

...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...

...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...

...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...

...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 ...

...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...

...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...

...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...

...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...