...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...

...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...

...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 ...

...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 ...

...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...

... 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...

...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 ...

.... 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...beginning your experiments. With CombiGEM and CRISPR platforms now being integrated, we look forward to the realization...

... 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...

... 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...

...double-stranded break. HDR copies the sequence from a repair template with flanking sequence homology for error-free...systems, and NHEJ will be favored even if a repair template is present. HDR also presents a cloning dilemma...’s become clear that the power of this editing platform lies in the diversity of nucleases and their applications...

...liquid media or solid media (on plates). In general, amplification on plates can help to ensure that diversity...library varies significantly in plasmid size). On plates, each transformed bacteria will be its own colony...

... to guides in the Broad’s Genomic Perturbation Platform Avana library (used by the Cancer Dependency Map...and Adam Brown from Broad's Genetic Perturbation Platform to design a new library that is agnostic to ancestry... are working with Broad's Genetic Perturbation Platform to modify the CRISpick algorithm to prevent guides...

...PE6b was very efficient for short, unstructured templates and significantly reduced undesired edits (indels...complex edits, such as long or highly structured templates. Cas9 variants PE6e–g often had even higher editing...potential to reduce negative side effects. This platform offers a promising approach for experimental therapies...

...general, technical replicates can be thought of as "plate controls". They are NOT independent and are typically...transfection of multiple separate wells (within the same plate) with purified plasmid from the same aliquot/preparation... the same source and were incubated on the same plate at the same time. Although the replicate control...