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...preferred platform before you start can save a lot of trouble in the long run.  Selecting the method Once you... Requires regular content production. Internet "trolls"   More resources on the Addgene blog:  Summer...  Summer SciComm Series: Cognitive LoadIntro to our Summer SciComm Series    ...

... and negative (gray) droplets. The no template control wells should have very few positive droplets. In...positive copy per microliter in the no template control wells. You’ll also notice a reduction in the number...assay, so cross contamination into a no template control (NTC) well is common. Here are some steps you can...

...provided a major toehold into understanding how genes control biology (a fact celebrated by the Nobel prize committee...transgenics Creating transgenic organisms by introducing foreign DNA into the genome is useful for a variety...into adults and explain how signaling pathways control this process. Additional zebrafish applications...

...advantages. An introduction to Gateway technology The Gateway cloning method, developed by Invitrogen, is an ...an in vitro version of the integration and excision recombination reactions that take place when lambda...

...of symptoms in animal models, Rett syndrome is a strong translational candidate for gene replacement therapy...therapy. In 2014, RSRT brought together a strong collaboration between two gene therapy labs (Brian Kaspar...neurological disease (Sinnamon et al., 2020). They introduced a mouse MECP2 guide RNA and a hyperactive version...

...laboratories and associated publications, but our quality control (QC) standards have evolved over the years to keep... ability to re-use or borrow existing code and control how the various pieces were connected, as this ... for Bioinformatics AnalysesAAV Vector Quality Control: Going the Extra Mile with NGS...

...degraders. But CRBN has been difficult to study in vitro, requiring either expressing it with additional ...codon-optimized for C. elegans and included artificial introns to improve expression. When using these constructs... an edited strain carrying Cas9 to allow the introduction of additional sgRNAs. Image reused from Neo ...

...a large number of requests for vectors in the “controls” category, which includes things like viral vectors...vectors in a variety of serotypes, including the retrograde AAVrg and the systemic capsid PHP.eB.   ...Patrick, M., Guerin, K., & Fan, M. (2023). Quality control for Adeno-associated viral vector production. Neuromethods...

...Laboratory Golden Gate Cloning videos made by the Nicola Patron group, one of the developers of the MoClo system...extracting and sequencing the plasmid, or both! We strongly encourage you to sequence the entire insert region...direction (which is not easy to see in colony PCR electrophoresis), is free of undesired mutations, and that ...

... for saturation mutagenesis, and they provide a strong proof of concept for further development. ...excision repair pathway. The team behind AYBEv3 introduced the first deaminase-free base editor when they... called gTBE and gCBE, with Addgene. gTBE can introduce T → C and T → G edits, though with limited selectivity...

...protein instead localizes to the cytoplasm? Or more troubling, what if a significant fraction of the fusion ...these compartments using CRISPR systems should strongly consider using moxFPs over standard FPs for the...Structural Biology. His interests include the quality control of secretory proteins in the Endoplasmic Reticulum...

...pGP-AAV-syn-jGCaMP8f-WPRE (AAV Retrograde) pGP-AAV-CAG-FLEX-jGCaMP8m-WPRE (AAV Retrograde) pGP-AAV-syn-FLEX-jGCaMP8m-WPRE...have Kd around 70 mM and 100 mM, respectively. Introducing mutations and using potassium-binding protein...

...epigenetic silencer Recombinant antibodies for marking astrocytes MagIC-Cryo-EM for sample enrichment and structure...determination By Mike Lacy Conventional cryogenic electron microscopy (cryo-EM) protocols typically require...glia-specific intermediate filament expressed in astrocytes throughout the brain, making it a popular target...

...transcriptional regulation, as long as they can control recombinase activity. For example, post-transcriptional...methods section of our paper (1). How might one troubleshoot GSRMs that don't work as expected? The successful...the parts used to build them. We recommend using strong, directional terminators and promoters with buffer...

...the number of integration copies can be tightly controlled by titrating the transposase to transposon ratios... levels of your gene of interest, as needed. Controlling the expression levels of your gene may be particularly...splinkerette-PCR method for the isolation and sequencing of retroviral insertion sites.,” Nat. Protoc., vol. 4, no. ...

...handed out at the event). Program 0 - 1:24 - Intro 1:24 - 29:31 - Reproducibility Overview - Jeffrey... Addgene as a source of sequenced and quality controlled plasmids and viral vectors. When working with... easier to learn new experimental techniques. Electronic lab notebooks also play a role here by making...

...CRISPR/Cas9 allows us to delete sequences and introduce mutations and epitopes with unprecedented ease...in 1-2 kilobases of additional sequence being introduced. Cre-mediated excision of the selective cassette... ML, Jorgensen EM (2018) SapTrap vectors for introducing point mutations with unc-119+ selection. https...

...and LIIbeta F 1-2 RNAi, which allows assembly of intron-spliced hpRNAs through Golden Gate cloning. Artificial...Identifying actively silenced plants Once you’ve introduced your RNAi into your organism, how do you identify...insertion of any target sequence, as well as essential control plasmids. Many thanks to our guest blogger Robert...

...substance (fluorophore) that has absorbed light or electromagnetic radiation at another wavelength (Heinrichs,...Heinrichs, 2009). When a fluorophore absorbs light, electrons are elevated from a ground state to an excited state...After a brief elevation in this excited state, the electron returns to its ground state by emitting a photon...

...happy cloning! This article was written by Susanna Stroik and Meghan Rego, with thanks to Rob Hurt for his...Plants Mediated by a Cas9 Gene Containing Multiple Introns.” Plant Communications, vol. 2, no. 2, Elsevier...Matthew S., et al. Degron-Based bioPROTACs for Controlling Signaling in CAR T Cells. Cold Spring Harbor ...