...gradients, practice making the iodixanol layers before doing your first purification. This helped me get a feel...to learn more about different ways to titer AAV. Do you have any AAV production tips or tricks? Please...multiplicity of infections (MOIs), to determine the optimal dose of AAV for my cells of interest. Every batch of ...

...interference), and CRISRPa (activation) libraries that do just that (Doench et al., 2016, Sanson et al., 2018... identifying strong hits in genetic screens, they do have several limitations. CRISPRko results in a complete...

...expression. To design a viral vector system that could do both, Addgene depositor Didier Trono and collaborators... lentiviral system (Figure 2). The HIV genes that do remain are very important for viral production: Gag...

...Gene X in the untreated cells—but can we be certain? Do we know whether the plasmid delivered into the cells...negative controls that produce a similar product, but do not target an endogenous gene in your experimental...

...of necessary data requirements and likely will not do so (calling it a “sisyphean task” given the endless...Firstly, we rely on many other labs to teach us how to do experiments and understand their data. Likewise, ...

... Michael Früh submission, which we have entitled "Do You See What I See?" I was bored this weekend while...

...quantification so that you can focus on what you want to do with your plasmid without spending time and resources...

...other aspects of molecular biology. “What I'd like to do with the plasmids from Addgene is have the students...

...expect given the excitement surrounding anything to do with CRISPR, our most popular post in July (with ...

...plasmids for replication and protein expression. So how do scientists recombine DNA? There are many methods ...so dissimilar from restriction enzyme cloning). To do this your DNA fragment must first be amplified with...move it into a series of plasmids that allow you to do many different molecular biology techniques (such...

...into 911E4 cells for virus production, as 293 cells do not contain E4. Adenovirus Packaging Methods Return... IRES-GFP pShuttle Class of transfer vectors that do not contain GFP Additional Vocabulary Term Definition...

...: Apply library Apply selection Most cells die or do not pass selection (in the case of a reporter) Sort... screen, the goal is to identify those cells that do not survive the selection mechanism. You’ll infect...

...wild-type and engineered variants of Cas nucleases. In doing so, they also created a modular toolkit for Cas-...

...all the excitement and speculation, we thought we’d do a little digging and assemble a rough guide to the...

...Asked Questions (FAQ) about γ-Retroviral Plasmids How do engineered γ-retroviruses differ from lentiviruses...promoter is no longer present in these constructs. Why do many γ-retroviral plasmids have similar names? The...

...Expression in Mammals AAV Titers: Where do they come from and what do they mean? A Match Made in Heaven: CRISPR...

...being published on our own topic. Only on occasion do we then have time to read about research from other...otherwise never learn from the valuable research we are doing. Browse the site, learn from others, share ideas...

... using site-directed mutagenesis of Spinach. In doing so, they identified the nucleotides that contribute...Just as vegetables come in a variety of colors, so do aptamers! Corn was recently described in 2017 (Song...

...editor. Cas13 enzymes are advantageous because they do not require a PAM, and RNA targeting is potentially...flanking sequence (PFS) adjacent to the target, but many do not, showing the flexibility of this system.  As ...

...array of base editing reporters that can help you do just that. Monitoring APOBEC and Cas9-mediated editing...tested their hypothesis with GFP fluorescence. To do this they generated a silent GFP construct, that ...