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...recombinase-dependent vectors, this can cause challenges for the experimental design as these vectors can exhibit spurious...studying how neurons are physically connected, we can begin to understand how the brain works. It’s kind...roll) neurons! Neurons are really cool because they can transmit information, for example, from one of the...looking at how neurons are physically connected we can begin to understand how the brain works. There are... (Fig. 2). Viruses as neuronal scalpels Viruses can be injected into the brain and efficiently deliver...finding a virus that works well for retrograde tracing can be difficult. The in vivo workhorse adeno associated...rabies) naturally exhibit retrograde transport and can therefore perform retrograde tracing, but often at...

...information so you can tap them for advice. As a future alum, these are people you can “cold call” and ask...someone you trust. A good coach can make a big difference, if you can find one. Create your own peer mentoring...if there are other ways they can help you grow. Mentoring relationships can last for an hour or for a year...First, I will describe some of the many ways you can approach finding someone to give you advice and guidance...the training or skills you want to learn. Mentors can be senior to you or peers you admire.  If you see...away We should all have a “posse” of advisors who can help us in different ways and in diverse areas. It... you present or see your work product in action) Can help you learn the local “ropes” and become an insider...

...information can be entered using our online form or by using a downloadable spreadsheet. Either choice can be ...pre-publication plasmids so they can be available when the paper is published. Plasmids can be linked to a peer-reviewed... Strain Addgene can distribute your bacterial strains if they: are E. coli that can be used under BSL1...utilize your plasmids (these can be protocols, GenBank files, etc). We can accept sequence files in any...plasmid cannot be propagated in DH5α. For plasmids with highly repetitive sequences (which can be prone...process. After you have logged in to your account, you can begin the deposit process by clicking "Submit Online... download the .xls file. For the online form you can submit "Published plasmids" or "Unpublished or pre-publication...

...2 nm, which can lead to higher resolution for super-resolution microscopy. Nanobodies can be labeled with... the above listed molecular techniques, but they can also be expressed in bacteria and extracted with ... to occur: A) Multiple polyclonal secondary IgGs can bind to different epitopes of a primary antibody....multiple labels (green circles in figure 2 below) can be attached to each antibody. But being big is a... target antigen and the fluorescent label, which can reduce the potential image resolution of super-resolution...recognize only one primary antibody. This process can be avoided direct antibody fluorophore conjugates...Reagent Pros Cons Antibodies Bivalent Polyclonal Can be labeled with multiple dyes Commercially available...

...), which can be fused to recombinant proteins for one-step purification with glutathione, can also protect...instability. Prokaryotes can also have a hard time folding eukaryotic proteins. You can get your bacteria to...ubiquitin-related modifier (SUMO) can help with folding and stabilization, as can maltose-binding protein (MBP...Overexpression can also lead to insolubility, and aggregated protein is not useful protein. MBP tags can help with...biologists who need to overexpress and purify proteins can face any number of technical challenges depending...antibodies available for these tags really are good and can be used for western blots, IP, and affinity purification...unlikely to affect function, His-tagged proteins can be purified using metal-affinity chromatography, ...

... of the seemingly simple actions we can take to help ourselves can be an appropriate first step. Check...You can work later (in the evening, or in a few years) when life responsibilities are less You can work... a charitable cause.  While I can’t make it “all better” for you, I can offer some tips and tactics that...-don’t list Prioritize! Some things can wait and some things can’t. I kiss my kids good night every evening.... Tell your mom you can’t make it to dinner this week. One swim lesson missed can mean a world of mental...soccer game. Tip: take turns going; that way you can have someone there clapping more often than not. ... your career will suffer. And, yes, your partner CAN take the kids to the doctor without your help. Your...

...enzymes that can convert certain chemicals into fragrances that they can smell, or enzymes that can create hydrogels...that students can easily sense, whether that’s bright fluorescent proteins that they can see, enzymes ...that they can see, smell, and touch. Along the way, we hope they’ll also see how fun biology can be! What...This is like taking an engine out of the car – you can still use the engine as a power source, but now you...replicating organism. Furthermore, cell free systems can be freeze-dried into pellets (FD-CF pellets), leaving...Most importantly, experiments for FD-CF reactions can be fun and engaging for students. In our BioBits ... hydrogels that they can touch. With BioBits, we want students to make the connection between the facts...

...acid tag (DYKDDDDK) that can be added multiple times (often 3X FLAG, but can be more!) to a protein’s ...immunofluorescence where an antibody can’t be used or that step can be skipped. It has the flexibility ...compatible, and doesn’t come with a high background can be a huge challenge. Epitope tags eliminate the need...the antibodies used to detect them, and how they can fix your antibody woes.   What are epitope tags?...possible, without the need for an antibody. Tags can enable the separation of detection of a mutant protein...when both are expressed in the same system. Tagging can also aid in protein purification! In this blog, though...s termini to increase detection. The tag can also be removed by treating the protein with enterokinase...

...neurons, you can see how it contributes to behavior or pathology, and by disabling neurons you can figure out...days. Addgene: Can you tell us a little more about this new two-color tool and what it can enable? Boyden... maybe we can get a handle on some of these mysteries. Another application is clinical; can we find ways...the engineering of its function." Once scientists can do that, they will not only understand how the brain...neuroengineering Addgene: For those who may not be experts, can you give me a brief history of optogenetics? How ... much, much, much more light-sensitive. Then you can use dim blue light and red light together. Understanding... some of the big problems that optogenetic tools can help us to address? Boyden: There are two concrete...

...system. The first two can serve as controls in a variety of experiments, while rtTA can be used to activate...that can be used to deliver fluorescent proteins to cells as determined by the serotype. You can use these...that you can use to map neuronal connections. Expanding the viral vector knowledge base: You can help! We...from select vectors in the repository yourself, you can now order ready-to-use virus directly from Addgene...many researchers we’ve met with have asked if we can help them share and distribute other reagents. Virus...while still ordering viral vectors. Virus production can be a frustrating process that requires a large investment...viral production and testing so that researchers can use our educational resources and experience to their...