...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...

...communicate outcomes to the rest of their team. Here's a template for project meetings. Team Meetings: For standing...members still have their voices heard. Here's a template for team meetings. One-on-Ones: Managers should... agenda template...

...concluded “Aspirin promotes autophagy, inhibits platelet aggregation, and potentiates the effect of PD-... UCSF Immunoprofiler - a human biopsy analysis platform focused on analysing immune measures and correlating...with tumor mutations and clinical responses. This platform has the potential to become an open-source pipeline...

...results? Or considered why DpnI will degrade your template DNA from a PCR reaction but not the newly synthesized...no methyltransferases in the PCR reaction). The template plasmid, on the other hand, should be derived ...with DpnI, only the non-mutated and methylated template is destroyed leaving behind a pool of mutated ...

... the cell with a mutant template and fooling the cell into using that template for repair. By using these...the two ends back together or using a homologous template provided by the sister chromosome. As cells take...

.... CombiGEM-CRISPR: The marriage of two simple platforms CRISPR has revolutionized how we decode the genome...regulation in biological systems. The CombiGEM platform provides a means to create barcoded gRNA libraries...Wong et al., 2016). With CombiGEM and CRISPR platforms now being integrated, we look forward to the realization...

... 2014). The second was an in vitro paper-based platform that allowed gene circuits to be deployed out ... had been working on advancing the paper-based platform toward a more field-ready format when the Zika...we say the timeline to develop our paper-based platform into clinically relevant diagnostics for the Zika...

... a fluorophore or tag. To utilize HDR, a donor template containing the desired sequence flank by homology...Nanoblades which carries a gRNA, Cas9, and a HDR donor template DNA can be generated by incubating Nanoblade particles...facilitated HDR with either a ssDNA or dsDNA donor template. In vivo editing of mouse zygotes and mice with...

...that grew on a selective plate versus the number that grew on a non-selective plate. Most bacteria tested...glmS gene in the recipient's genome. Antibiotic plates lacking DAP select against donor bacteria and for...