... to glance at as you work. Draw your plate maps onto your plates, alongside your physical copy. If you...reminders on a sticky note   Visual Planning  Plate maps, pre-labeling tubes, and even drawing out experimental...example, setting up your pipette tips to match your plate layout can help you track samples as you add them...

...desired mutation, then use those to PCR amplify the template plasmid. The parental, nonmutated DNA is digested...nick repair. This method can work well, but poor template digestion can result in a high number of background...mutations, to amplify both strands of a plasmid template. In addition to containing the mutations, the ...After being treated with DpnI to digest away the template plasmid, these PCR products are directly transformed...

...samples through our lab using barcoded tubes and plates. The critical pieces of information for a given...100 µl of glycerol stock to each of 2 96-well microplates (identical copies, see image above). We then ... database of over 2000 primers. Two 96-well PCR plates are prepared with the primer stocks for forward...shipped along with their corresponding glycerol plate on dry ice to the sequencing provider. Each sequencing...

...each gene, we needed to design guide RNAs, donor templates and sequencing primers for quality control. At... taken up to a month to design a single 96-well plate for HDR that satisfy all requirements. To alleviate...guides by CFD score, and it mutates HDR donor templates to avoid re-cutting when needed. It gives you ...the standard information that goes into a 96-well plate and comes out of it. Biotech companies are known...

...genes. For this you would need to provide a DNA template to repair the double strand break via homology...Cas9/sgRNA and one vector for your specific DNA template. Using this two vectors system, Manuel Gonçalves...This technology therefore represents a potential platform that could be realized via simple switching of...tools can also be used to deliver Cas9 and DNA templates in somatic cells of adult animals in order to ...

...homologous recombination using a repair template. These templates can include loxP sites to create conditional...recombination. The concept is simple: deliver a template with a modified version of the targeted sequence...sequence to the cell which will recombine the template with the endogenous gene. Here, we'll describe the ...

... from an LB plate. In my hands, a 5 mL starter culture inoculated from a colony on a plate will reach ..., check out Harrison et al., 2006. Tips: Don’t plate seeds too densely during selection: this will increase...difficult. Don’t leave your plants on selective plates for too long: even resistant plants will be affected...

... to copy and modify templates (e.g. http://openwetware.org/wiki/Protocols/Template). Nature Protocol Exchange... up after trying out the new protocols.io beta platform for sharing, annotating, and storing life science...molecular cloning protocols, including making LB agar plates, restriction digest of plasmid DNA, and bacterial...

...strategy for increasing the size of the AAV donor template for CRISPR-mediated homology directed repairs ...homologous recombination approach delivers large donor templates using two sequential homology directed repairs... the gRNA target site that mediated the donor template’s integration so that the site is reconstituted...which results in the integration of a larger donor template.  Drawbacks: Transgene expression varies with...

...broth and on the agar plates. Mycoplasma colonies are easily visible on the agar plates. This method is capable...detection, DNA staining, fluorescent tagging, agar plating, etc. Be wise to choose any of the techniques listed... (Uphoff and Drexler, 2011). Culturing on agar plates/broth is considered the ‘gold standard’ for detecting...

...in 16 wells of a 96-well plate, allowing 6 conditions to be screened per plate. The results indicated that... pools were prepared consisting of one target (template) strand and four upstream probes and four down-stream...strand covering all 16 possible NN pairs in the template. The probes were designed such that each possible...

...LinkedIn. We are also exploring other social media platforms with active scientific communities and welcome...functionality and features that previously made the platform a mainstay of conversation in the scientific community...considering a gradual move off X as a response to major platform changes, such as limiting access and increased...

...cloning have been utilized to create recombination template vectors, to perform multiple site mutagenesis ...Engineering TALENs containing variable-repeats using Platinum Gate system Interested in optimizing TALEN assembly... single-strand annealing (SSA) assay. This new Platinum Gate TALEN Kit utilizes a 4-module assembly system... plasmids (ex. p1HD, p1NG, p1NI & p1NN) in the Platinum Gate system contains an identical variable repeat...reports. For additional information on using the Platinum Gate TALEN Kit, please see the Yamamoto lab’s ...

...incubation step and go straight to plating if your transformed cells are being plated on agar with ampicillin selection...competent cells, don’t forget to do a negative control (plate your cells without adding plasmid) to make sure ...

...Method Notes Quantification of a process Plate-based assays (plate reader) Detect the average amount of ...fluorescence across a population of cells (i.e. in a plate well). Use to track a process (e.g. enzyme activity...

...enough time to completely digest the template DNA. Note—templates must have been isolated from methylation...sensitive restriction enzyme DpnI to remove the templates. Digestion products are then transformed directly...

...Biology 2018 Pubmed PMID 29379139. The “Riboglow” platform for visualizing RNA production and localization...cobalamin binding riboswitches used in the Riboglow platform at Addgene. Braselmann E, et al. Nature Chemical...a new imaging-based bacterial colony screening platform, Oliver Griesbeck’s lab set out to engineer a ...

...fields of biology and enable the conversion of RNA templates to DNA but suffer from several important shortcomings...probability of the polymerase continuing to copy the template rather than dissociating).  MarathonRT, isolated...structured HCV genome (~9.6 kb of highly structured template!). MarathonRT also faithfully transcribes RNA ...

...repair template plasmid to create fluorescent and 3xFLAG-tag fusion proteins. The repair template and Cas9...worms have laid eggs, hygromycin is added to the plates to select for candidate knock-ins. These candidate...

...can be counteracted by growing the cells on solid plates, where each transformant forms its own colony, ...less of a concern, but Addgene still recommends plating the transformation mixture on solid media. The ...The colonies are then scraped off the plates and pelleted for DNA extraction. Prepare library product for...