...template with the endogenous gene. Here, we'll describe the techniques and the plasmids used to inactivate...Conditional knockouts The methods and plasmids described in the preceding sections are simple ways of knocking...knockout, researchers often use the Cre/Lox system described earlier. In this case, you design your targeting...

...photostable than fluorescent proteins, making them ideal for single-molecule fluorescence techniques. After...used to measure signal along the dendrite. D) Fluorescence signal as a function of the distance between...

...proteasome for degradation. Like several other tags described here, the dTAG uses a small molecule, a tag, and...for visualization of proteins of interest. A fluorescent Halo ligand can be added to media, buffer, etc...there are antibodies to it and ways to achieve fluorescence with it. AID The auxin-inducible degron (AID...

...following cloning methodologies have been extensively described in other articles; so if you are interested in... to assemble up to 6 different fragments in a prescribed order. These fragments can be pooled in one tube...homology. You might also consider FastCloning, first described by Chaokun Li and colleagues. Here you simply ...

..., if your protein is fused with a fluorescent protein), fluorescence microscopy can theoretically be used...Addgene.org Check Out Our Protocols Pages Browse Fluorescent Protein Plasmids Find Mammalian Expression Vectors...

...organism carrying a synthetic gene drive were to escape the laboratory, the drive could eventually spread...protocols, they can reduce the risk of accidental escape to a negligible level. One major reason we chose...performing experiments in a geographic area where escaped organisms won't be able to find mates2. For example...

...colorimetric ladder, visible to the naked eye; a fluorescent ladder compatible with your imaging system; or...having to re-run the western.  Remember that chemilumescent methods, like horseradish peroxidase protein...Underexposed Increase exposure time Chemilumenescence: use a more sensitive exposure kit Too...Overexposed blot Shorter exposure time Chemilumenescence: use a less sensitive exposure kit No ...

...looking at your resume while they wrote the job description. You check off all the boxes - you have the desired... gaps in scientists' transferrable skills. In describing the limitations of the study, the authors noted...over email. Examine your current and past job descriptions. Scan through all the things for which you are...

...are easy to trace and not lost in a jumble of fluorescent neurons. A simple but flawed approach to multi-color...constitutively express a red, green, or blue (RGB) fluorescent protein (4a-c). When high doses of this viral...morphology studies when used as part of a two part fluorescent tag-then-express system that allows for controlled...

...worked extensively with enChIP systems. Can you describe this technology and its advantages? Fujii: In ...enChIP system using CRISPR technology. Can you describe this technology and some of its potential applications... any other technologies that you would like to describe?  Fujii: I believe that "Biology is Method", i.e...

...used in research, and data repositories. Below I describe how each of these kinds of connectivity facilitates...deposits will be made. The Dataverse integration is described in this video and the Figshare integration in ...from RSpace documents to external data sources (described above) further enhances the value the repository...

...every amino acid I’d like to encode,” but, as described above, not every region of a protein should necessarily...rate dependence of transfer RNA abundance in Escherichia coli." The EMBO journal 9.13 (1990): 4359-4366...rate of full‐length human proteins expressed in Escherichia coli." Protein Science 19.7 (2010): 1312-1326...

... have been made since prime editing was first described in Anzalone et al., 2019, including modifications...Because they are negative-going sensors (with fluorescent signal at baseline voltage), the investigators...closely-packed labeled cells. Plus, Voltron2’s fluorescence is tunable by changing the dye that becomes ...

...areas associated) compared to an animal running to escape a threat. Locomotion can thus be used as a readout...behaviors are possible: active swimming in attempts to escape, or passive floating on the water surface. The ...that were not (Hare et al., 2019). The research described here is only a small selection of the increasing...

... through Addgene. In addition to the plasmids described above, Addgene carries several useful CRISPR toolkits..., Cao Q, Zhang Z, Ding Z, Wang Y, Wang X (2019) Rescue of a plant cytorhabdovirus as versatile expression....2969 Waltz E (2016) Gene-edited CRISPR mushroom escapes US regulation. Nature 532:293–293 . https://doi.org...

...genome editing with CRISPR/Cas9 in human cells was described in 2013 (Cong et al, Mai et al) we were very excited...thinking further about the distance relationship described above, we realized that, in addition to increasing... and neurovascular diseases. He worked on the described project while he was a Postdoc in Marc Tessier-Lavigne...

...promoters can be used in the repressible manner described above, as well as the inducible manner detailed...30 years since the original Tet-Off system was described by Gossen and Bujard, both the promoter and transactivator...cells, and can be used in the repressible manner described above, as well as the inducible manner detailed...

...for and how can you do it well? Below, I briefly describe some tips for a successful postdoc fellowship.... there a tractable question or are you simply describing a phenomenon? Exploration can be important, but...in the Endoplasmic Reticulum, optimization of fluorescent proteins,  and live cell imaging approaches. ...

... been described in this blog. If your data does not meet the assumptions for the tests described above...error rate than the multiple comparison tests described above. Violated assumptions and statistics Statistical...

... community is generating a massive amount of descriptive transcriptomics data. Custom CRISPR screens designed...videos and text on the website for more detailed descriptions of how to use the software. Here is a brief ...sequences, and more. Output_Compact: A text file that describes which genes were included, and how many gRNAs ...