...connectivity to other tools used in research; and (3) connectivity to open source data repositories. To...RSpace is integrated with most of the tools noted above, and has a modern API. Integrations with additional...into RSpace. The kind of informal conversations about research that can take place in Slack normally never...RSpace documents to external data sources (described above) further enhances the value the repository integrations...Minisymposium on Reproducibility Read all blog posts about scientific sharing Resources on Addgene.org Find...

...Orthogonal Type VI-A CRISPR-Cas Enzymes." Mol Cell. 66(3) (2017):373-83. PubMed PMID: 28475872. Find plasmids...CRISPR Featured Topic Page Learn about RNA Targeting with Cas13a Learn about CRISPR DNA Base Editing Resources...and sequences that can be edited.   As discussed above, the ability to edit RNA has multiple advantages...

...sample to a mastermix. At Addgene, we typically load 3 dilutions onto our ddPCR plate. As the titer is unknown... droplet generator, you will have to be careful about transferring your droplets from the droplet generator...Additional resources on the Addgene blog Read more about polymerase chain reaction Browse our blog posts ...collection of viral vector related protocols Learn more about the Addgene viral service ...

...www.businessinsider.com/how-to-get-the-most-out-of-job-fairs-2014-3 https://www.forbes.com/sites/neilpatel/2015/05/21/...or postdoc forever, and you need to be thinking about what comes next. To that end, one of the most important...personal connection; the importance of which will be elaborated on later. As for what job fairs to consider, ...the specific position he or she originally called about, you can keep in touch and reach back out when you...

...often be seen by sinking of the tissue (see Figure 3). This method is probably the most scalable method...Generating a transgenic plant takes even longer, is more labor intensive, inefficient, and often not reproducible... region is transferred into the plant. In the laboratory we can utilize this natural process by replacing...system. The vectors mentioned in this blog post above are ready to use vectors, but with the MoClo system...

...addiction module" regulated by guanosine [corrected] 3', 5'-bispyrophosphate: a model for programmed bacterial... more about CcdB, the toxic key to efficient cloning Read all Plasmids 101 blog posts Read about knockout...with the other toxin-antitoxin systems described above, one part of the pair is more stable than the other...

... sequence complementary to the mRNA sequence. The 3’ end of  the tRNA binds the amino acid that corresponds...usage bias Read more about plasmids in our Plasmids 101 blog series Learn more about trends in synthetic... out Addgene's Molecular Biology Reference Read about genetic code expansion Browse our plasmid collection...

... the subsequent bbmerge process.     Fig. 3: How we get bbdone with our bbduk process.    ...much information as possible from depositing laboratories and associated publications, but our quality...the process. Another design choice we appreciated about Nextflow was how effortless it was for us to switch...interface. In our next post, we’ll share our story about how we used Nextflow Tower to augment our pipeline...

...specifically to the blue triangle protein but not the other. 3) The beads are collected by centrifugation or a magnet...proteins or nucleic acids. Read on to learn more about this versatile technique.  Immunoprecipitation ...proteins in an IP are not denatured (learn more about antibodies here!) When using Protein A/G to immobilize...eluted by boiling the beads in SDS. (To learn more about Western blotting check out our blog post The Basics...

...darker than full vector particles.     Figure 3: Electron micrograph of AAV vector preparation shows... before being distributed to customers. Details about our production protocols, titering methods, and ...either in-house by Addgene scientists or through collaboration with viral vector manufacturing facilities, ...generated from lentiviral preparations (described above) that are subject to a concentration step prior ...

...immunoprecipitation (enChIP)." Scientific reports 3 (2013): 3171. PubMed PMID: 24201379. PubMed Central...specific genomic region In 2013, the Hodaka Fujii Laboratory first described a method to purify a specific...additional research conducted in the Hodaka Fujii Laboratory, please also see our interview with Dr. Fujii...

...–798. https://doi.org/10.1016/s1097-2765(02)00496-3 Kerppola TK (2006) Design and implementation of bimolecular... FP fragments and, preferably, some information about the structures of the tested PIPs. Many split FPs...neurodegeneration. He now spends his time learning and writing about science whenever he's not off the grid.   References...

...PMID: 25599551. PubMed Central PMCID: PMC4344395. 3. Lavis, Luke D., and Ronald T. Raines. "Bright ideas... the frontiers of biological imaging.  *Worried about your budget? Email [email protected] Resources on the Addgene Blog Learn About Other Protein Tags Catch Up on Your Fluorescent ...

...plasmid fingerprinting", where you cut the plasmid into 3-8 pieces such that all (or most) fragments are small...insert, but significantly off center (ideally around 1/3 of the way from one end), and also cuts in the backbone...verification, such as DNA sequencing . In the example above, digestion with enzyme RE1 will linearize the 6200bp...that you get double confirmation. In the example above, digestion with either RE3 or RE4 will give a very...

....   The Dodt and Frank Bradke laboratories later developed the 3DISCO (3-dimensional imaging of solvent...

...the membrane is cut so each lane is its own piece (3). After incubation, blocking, and washing, the membrane...protein concentration decreases).     Figure 3: Testing the antibody’s dynamic range. A) The protein...re in the right place! In this post, we’ll talk about western blot optimization and troubleshooting.   ...can be informative, so spend some time thinking about which type of gradient can help answer your particular...overexpression. Before starting a western blot, think about how you’ll interpret the data and what level of ...

...anneal to the regions upstream (5’) and downstream (3’) of the DNA segment to be amplified. When these reagents... increase the time of the denaturing. Your 5’ and 3’ primers should be designed to have similar melting...Reverse Primers: Hybridize and are complementary to the 3’ ends of the sense and anti-sense strands of the template...manufacturer’s instructions for specific instructions about extension time and temperatures. Initial Denaturation...

... host told me and wished I had been more careful. 3. Tell a story We are biologically programmed to listen...interesting content Talk about something you know a lot about and that you care about. Don’t try to present...engaging and memorable. If you read my story above about the presentation faux pas, you are more likely...the room, so it is a good idea to start thinking about public speaking early and often. Two of my jobs ...dynamite presentation?  To that end, let’s chat about giving science talks and how to make them serve ...other people give great talks We are not talking about lab meeting here. Lifeless presentation of experimental...You want to give the audience something to think about, argue with, or learn from. Some of the best talks...

...write our AMA submission to avoid rousing the trolls. 3. The AMA process was simple and well organized. You...not too worried about selling yourself and really do just want to get information about yourself and your...where users can participate in forum discussions about a wide variety of topics. Individual forums are ...where users can ask designated scientists anything about a particular topic. Given Joanne’s expertise in... is well moderated. One of our original worries about sharing science advice on Reddit was that we’d be...answering questions. r/Science has pretty strict rules about not being too sales-y in posts which falls in line...any scientists to refer to if they have questions about science careers, women in STEM, work-life negotiation...

... remember that antibodies recognize very specific 3-dimensional epitopes. Different sample processing ... Learn about research applications of antibodies in Introduction to Antibodies Learn more about fluorescence...Stay tuned for a future blog post to learn more about these techniques!  Direct vs. indirect immunofluorescence...different approaches to be familiar with when talking about IF, direct and indirect. For direct IF, researchers...immunofluorescence procedure with key considerations about each step. Image created with BioRender.com ...of protocol you will need to use. (To learn more about antibody validation, this commentary by Uhlen, et...Fluorescence Microscopy Techniques is Best for Me? Read about another common antibody application in The Basics...