...cut site location Position your CRISPR cut site as close as possible to the knock-in location. HDR depends...sequence to be incorporated at a precise genomic location, with little to no room for error. It’s no surprise...nutrients in their media, they will not undergo DNA replication, so make sure you are keeping an eye on those...

...Dependency Map cancer cell lines (left). Heatmap indicating the breakdown of ancestry associated dependencies...machinery where to cut.      Fig. 2: Heatmap indicating loci where SNPs reside on sgRNA target sequences...Understanding the impact Fig. 3: Histograms indicating the frequency (x-axis) which SNPs map to thetargeting...same reference genomes and libraries. Further complicating the issue is the long and ugly history of reporting...

... and have been adapted for a wide variety of applications - as mutagenesis tools for genetic studies (... The process also creates a 5-bp target-site duplication (TSD), a hallmark characteristic of the Tn7-family...the new streamlined VchINT system was shown to catalyze insertion of mini-transposons up to 10 kb in size...at one or multiple desired genomic loci. Many applications of these systems have already been demonstrated...

...Subsequent advances in fluorescent proteins have replicated many of the properties once exclusive to small-molecules...phosphopantetheinyltransferase) – These enzymes catalyze the covalent attachment of a fluorophore ligand...labels for fixed-cell imaging, not for live-cell applications. Thus, these fluorophores are often bulky and...the Addgene Blog Learn About Other Protein Tags Catch Up on Your Fluorescent Protein Background Which ...

... can often be accounted for using appropriate replicates and controls (see Tips below), and by validating...two-hybrid screening Minimizing false positives: Run replicates of the experiment to reduce the likelihood of...expression and correct folding, post-translational modification, protein degradation, access to the nucleus ...co-expression of the enzyme responsible for the modification in the assay host strain can help. In order...

...either one at a time or as a batch of hundreds, and indicate the guide RNA sequence you used. The ICE tool ...are no parameters that need optimizing and no complicated steps to learn. For increased flexibility and...checked circle. Typically, a yellow check mark indicates that a particular parameter was automatically ...support team at Synthego. Her work centers around educating and supporting novice to expert-level researchers...

... gives you a single level of control without modification - either Cre is there or it’s not. Cre-mediated... sites can lead to excision, inversion, and translocation of DNA in a similar fashion as between loxP ...37 °C). Smart molecular evolution led to the identification of FLPe which has a temperature optimum of ...-LDB domains of the fusions allowing Cre/FLP translocate to the nucleus where they exert their recombinase...

...attP sites of the donor vector. This reaction is catalyzed by the BP Clonase enzyme mix and generates the...sites of the destination vector. This reaction is catalyzed by the LR Clonase enzyme mix. As a result, an ...carry different antibiotic resistance markers (indicated here by plasmid color), allowing you to easily...is simple -  no restriction, ligation or gel purification steps are required! Multiple fragment cloning...

...cancer. These later stages of a cancer diagnosis indicate, for Stage 3, that the cancer is large in size...size and may have spread while Stage 4 indicates that the cancer has spread from where it originated to ... dire prognosis for mesothelioma patients is to catch the disease earlier. New techniques are currently... working to fight the cancer.  Beyond early modifications to standard treatments and early detection, ...

...Marker-independent identification of glioma-initiating cells. The International Cell Line Authentication Committee...propagation of the line, and can include assays for authentication and contamination) and cell lines can become...let alone the human cost. Here are just a few publications which have since been retracted due to cell ...

...resistance gene and the origin of replication are necessary to replicate plasmids and make sure the cells...length version is strong with high expression. Truncated promoters are constitutive with lower expression...metabolic operon Inducible by arabinose and repressed catabolite repression in the presence of glucose or by competitive...

... the planning section below, it’s important to dedicate specific amounts of time to specific tasks beforehand...instance, if I don’t have draft blog post ready for publication for tomorrow, I’d better upload that post from...handle doing a particularly annoying task and then dedicating that much time to it each day. Once you get over...Student Community Learn about Careers in Science Communication Resources at Addgene.org Check out Careers...

...Creating great educational science videos is tough. Is the content accurate? Are we conveying complex... had set up involved multiple people in tight locations. Conference rooms for on-screen presenters! Labs...wasn’t going to work for the majority of our educational videos. In addition, it’s really asking a lot...receiver, to beam the camera’s image to my remote location, live. After doing a bit of looking, I found an...

.... After identifying all the major equipment, categorize all of your minor tools, like pipets, or mini-centrifuges...titles in the regions where the host institute is located. For example, salaries in Kansas versus salaries... most start-up P.I.’s. It doesn’t have to be complicated, or difficult to create this list. Start BEFORE...discounts, collaborations, access to internal award applications, etc. If this is the institute that can help...

...of farnesylated derivatives enables labeling of delicate axonal and dendritic processes not previously ...Between Brainbow Systems   Brainbow variants and applications beyond the mouse brain In addition to improving...temporal control without the need for germline modification, and enables Brainbow to be used in a variety...technique will provide important insights into the complicated physical organization of the brain and other ...

...protein is (localization) and how much there is (quantification). But what if you’ve got several interesting...primary antibodies that have been validated in IF applications (find IHC validated or ICC validated antibodies...informative image-based figures for scientific publications. PLOS Biology, 19(3), e3001161. https://doi....Immunocytochemistry Protocols for other antibody applications...

...vessel barriers. It is especially popular for applications targeting liver tissue. In addition, it has ...the blood-brain barrier, making it useful for applications targeting the CNS (Issa et al., 2023). Additionally...treatment, which can pose problems for gene therapy applications. Cross-reactivity of AAV serotypes varies by.... A. (2023). Various AAV Serotypes and Their Applications in Gene Therapy: An Overview. Cells, 12(5), ...

...at the Institute of Cancer Research in Sutton, located on the edge of London to gain relevant experience...recession. As each advertised job received many applications, employers were requesting that only scientists...commercial company. In my particular situation, communication between departments was sometimes difficult ... order to have meetings between teams. This communication is vital because otherwise I’d lose touch with...

...resource for anyone looking to use antibody-based applications in their research.  Like all our eBooks, the...through the process. If you’re confident in your application and protocols, but can’t quite seem to find an...for Your Experiment.” Whatever your antibody application question, the Antibody 101 eBook is a great place...

...light path. Each hole corresponds to a different location in the sample. As more than one hole is illuminated...questions will place each sample into one of four categories that are each suited to different types of modern...as spinning disk confocal. However, due to the scattering nature of biological tissue, imaging more than... allow for multi-view imaging can image thick scattering samples in their entirety, something that is ...