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...that can be harvested from the surrounding media. Nonetheless, there are a variety of ways you can optimize...increase the AAV concentration. In this flowchart, you can see the dwindling proportion of AAV particles that...their way to the nucleus where their gene cargoes can be expressed. The bottom line is that a lot of AAV...for this function is not yet fully understood, we can imagine that this transport is probably not 100% ...to drop off drastically with lower titers. As you can see in figure 2, the expression from the 5x1011 titer...considerations If you have high quality AAV, you can address the issue of low titer by concentrating your...AAV production. We’ve listed some simple steps you can take below but you should read our AAV Production...

...everything you do in the form of a protocol, and you can add in personalized steps like “check drawers for... next to each step as you complete it. Yes, this can be incredibly tedious, but it also works, so do it...organization of your workspace and your workflow can have a huge impact. Test different approaches, like... up your pipette tips to match your plate layout can help you track samples as you add them. Moving your...from your left side to your right as you use them can help you remember what's been added and what hasn't...hasn't. Developing simple habits like these can consistently reduce errors as you work, and the possibilities... If you need to focus, a large set of headphones can act as a Do Not Disturb sign, even if the headphones...

...research practices (QRPs) can quickly lead us to find statistically significant results that are not robust...difficult to do?  There are four reasons that research can be difficult to replicate. Unclear methods Until ...false negatives in replications are unavoidable, but can be minimized with high powered research designs. ...population, new genetic variation, or different handling can all be important.  If we eliminate the problems above... out the same as an original research finding we can be more certain that some important, previously unknown...other materials. The use of reporting guidelines can also help remind us to include all of the essential...before conducting the experiment (note: registrations can be embargoed while the research is ongoing, but all...

...occasional errors. That can be advantageous if you are trying to knock out a gene. One can also exploit homology-directed...people, we can use this kit to knock in those mutations. Once that's done in stem cells, we can differentiate...: For people unfamiliar with this new TALEN kit, can you describe it for us? Musunuru: TALENs are a type...type of genome editing tool with which one can introduce changes into the genomes of mammalian or other...is that the DNA-binding domain is modifiable and can be programmed to bind to essentially any DNA sequence...template for repair. By using these two scenarios, you can either knock out a gene or knock in a mutation. Addgene...more of an apples-to-oranges comparison. Addgene: Can you compare the TALEN kit to CRISPR/Cas9? When would...

...experiments, just about anything can disrupt your bench work and sometimes no culprit can be found…. In our lab ...experiments in Brazil can have the same issues as one doing experiments in France. We can learn with each other... knows that there are many ways your experiments can go wrong. Whether it be a bad fridge freezing your...discovered. As we started building, we thought – how can we make the connection between LEGOs and science ...differently than we do day to day. We discovered that it can be good for you and your experiments to think outside...Additional Resources on the Addgene Blog Learn How You Can Use Video to Share Your Research 5 Reasons to Use...

... aimed at a peer audience.  Note that lay people can and do consume scientific communication (it is, after...how one moves from layperson to expert) and peers can and do consume science communication.  Formal education... is provided, typically ad hoc, for learners who can choose to engage or disengage with a very high level... is field-specific terminology or language. This can refer both to terms that are only used in the field...non-jargon in the context of disease testing, it can still be considered jargon in most other uses. Many...that will amplify a sequence only if present.” You can see how you could end up in a very confusing situation...communications project without an audience?  An audience can be defined by any number of characteristics. Common...

...availability. An antibody can only bind to a target, or epitope, that it can access. If the epitope is...antibodies, which only recognize one epitope. (It can sometimes be an issue with polyclonal antibodies,...used as a marker for flow cytometry, for example, can only be recognized by antibodies that bind to the...a cell membrane.  Picking the Right Antibody  It can get even more complex if you’re looking for proteins...to translocate or fold in certain conditions? It can help to visualize an abstract protein during your...change depending on the conditions and the assay can help you avoid a great deal of frustration when designing...designing and troubleshooting immunoimaging assays. It can also elucidate information about what a given protein...

...gene they control. Promoters can be organism or tissue type-specific, which can help you restrict the expression...palindromic nature, and high GC content. This instability can cause partial loss of an ITR during routine plasmid...recombination-deficient bacterial strains, such as Stbl3s. ITR loss can be detected by restriction digest using enzymes, ...companies now offer Sanger sequencing conditions which can read through this challenging sequence. Transgene...map, its sequence is located between the ITRs. It can be anything you like as long as it’s ~4.4 kb or smaller...transgene to a specific species or location. Promoters can also drive different levels of expression. For example.... Lox sites are directional 34 bp sequences that can flank the transgene and are located between the ITRs...

...for viewing, editing or making plasmid maps, but can also analyze and annotate any DNA sequence. SnapGene... also has a suite of molecular biology tools and can make plasmid maps. Free for academic users. Serial...be “cleaned up” or changed in some way before you can analyze them. These tools help with that.  DNA sequence...Cloning ligation reaction calculators These links can help you calculate how much insert and backbone you...Calculator: This tool works for any cloning method. You can select from five different insert:backbone ratios...calculators Lab math calculators These calculators can help you with common calculations performed in the...iPhone Reagent and Data Repositories These links can help you find experimental and/or validation data...

...publishing posts can be addicting. Even more satisfying, because Addgene is a non-profit, we can focus more ...but instead can involve walking alongside it and giving it a high five. While screaming can be therapeutic...visit scientists in person to figure out ways we can make Addgene better and make scientists’ lives easier...many young scientists are led to believe that they can all end up in academic positions (I’ve even blogged...plasmid deposits, and trying to learn ways Addgene can improve in its mission to make research easier. I...outskirts, and entirely outside of academia. These roles can include positions, like mine, that help support academia...