...from Addgene 1. Location, location, location! What do buying real estate and using AAVs have in common?...each serotype can transduce multiple tissues. So how do you choose the right serotype? Keep asking questions...injection. Examining serotypes 1, 8 and 9, not only do they undergo retrograde and anterograde transport... transport can be reduced by injecting a smaller dose of AAV and using a serotype with weak CNS penetrance...

...based gene editing. But, how do you decide which tools to use, and what do you need to know about them ...similar to what the endogenous binding partner would do. This allows precise control over when and where ...

...Genome-Wide Screening What Are Pooled Libraries? How Do You Use a Library? Cas9 Fusion Tools Activation/Repression...to G (or T to C) change. Adenosine DNA deaminases do not exist in nature, but have been created by directed...have been developed for inhibition using dCas9. How Do You Use a CRISPR Library? CRISPR libraries from Addgene...alterations, such as small mutations or inversions, than do large deletions generated by Cas9 systems. Cas3 must..., Cogan, J. Z., Replogle, J. M., Adriaens, C., Ramadoss, G. N., Shi, Q., Hung, K. L., Samelson, A. J.,...

...non-coding RNA, cutting most anywhere within the RNA will do. Similarly, if you want to knock out a gene, a frameshift...introduced anywhere within the early coding region will do the job. What matters for these experiments is cutting...

... . https://doi.org/10.1038/srep31730 Tsuruta LR, dos ML, Moro AM (2018) Display Technologies for the Selection... 2015a). With molecularly defined antibodies that do not change over time, scientists will know exactly...Harbor Laboratory Publications, New York. Helma J, Cardoso MC, Muyldermans S, Leonhardt H (2015) Nanobodies...

...tool for gene delivery, but it has a drawback: how do you ensure your gene goes where you want it to? Knowing...approach to enhance specificity in retinal cells. To do this, the group developed a new strategy for finding...

...sgRNA. What if I want to work with gene drives? Then do so! It's a tremendously promising field that could...degree of confinement are those of the author and do not represent a formal stance taken by Addgene or...

...everyone gets involved. So join the conversation! Do you have a question about culturing iPS cells? Confused...

...detection range of your protein. The most common way to do this is via a Bradford assay or BCA assay. Be sure...preserve specific protein modifications, we recommend doing a little more research into gel chemistry and/or...wide dynamic range for fluorescent proteins. They do tend to have increased background with higher exposure...your blot to calculate the amount in your sample. (Do ensure your blot is optimized so your standard curve...

...our favorite posts from 2022! Wow! Our depositors do some really incredible science - I hope you've enjoyed...

...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...

...fluorescence when excited with ultraviolet (UV) light, they do so via aromatic amino acids found in all proteins...photons excite electrons of a fluorophore, they can do so at a range of wavelengths, rather than at just...1, emission spectra of neighboring colors can and do overlap. For example, APC overlaps noticeably with...

... a part of a community that recognizes doing science (and doing it ethically) runs hand in hand with policy...views represented below are those of the writers and do not necessarily represent the views of Addgene. ...

...biochemistry researcher at New England Biolabs. When do you need a high fidelity ligase—and when is an alternative...base pairing through minor grove contacts, and thus do not read specific base sequences, but are sensitive...differential is not known for certain, but may have to do with the slight melting of the 5 ́-terminus during...

... = 500,000 colonies.  Pro tip! Since you probably do not want to individually count 500,000 colonies, ...did not result in significant recombination. If you do see recombination, we recommend ensuring the cells...

...use But exactly what kind of editing are people doing with all these CRISPR plasmids (and libraries)? ...scientists use CRISPR technology? We think so — and so do our requestors! CRISPR plasmids in the Addgene repository...

...newer, such as the relatively recent resurgence of “Do-It-Yourself Science” described in Chapter 3, and ...never inherently bad or evil. However, it is easy to do sloppy science, misrepresent science, or misuse science...

...Do you ever wonder about the origins of some of the common techniques or tools you use in the lab? Take... to be less sensitive to sequence context.    So, do Schüchner, et al.’s results mean the end of 9E10’...

...guanines). This limits the use of ABEs as DNA sites that do not contain a suitable PAM cannot be edited efficiently...While base editors are precise molecular tools, they do have undesired off-target editing effects. ABEs have...