...grants. What is a grant professional?  In the past 5-10 years, the Grants Profession has become just that...on becoming grant writers. When we asked others about how they became grant professionals, mid- to late-career...late-career grant professionals have similar stories about a time when their passion for an organization or... logic models, established partnerships and collaborations, and commitments to and proof of compliance...grants. It is no surprise that the U.S. Bureau of Labor and Statistics shows that employment of grant writers... skills & passion  Skills such as research, collaboration across multiple disciplines, writing, project...development strategy builds on the steps listed above and formalizes your development. You may consider...

... Cells HEK-293T Sf9 Time to collection 2–4 days 5–10 days Purification Ultracentrifugation (AAV); ... widely available, so you’ll only have to worry about cloning your transfer plasmid. The transfer plasmids...typical insect cell preparations, which we talk about below. Processing The viral supernatant is typically...available, so the amount of cloning required is about the same between insect and mammalian systems. Baculovirus...dead cells and cellular debris. While adding time (about three to four days), this second round provides ... can skip the first round of production, saving about a week of work. An important note is that this is...

...control of the nervous system. Nature Neuroscience, 19(5), 756–761. https://doi.org/10.1038/nn.4265.  Additional...our tissues unimpeded. Andrew and Maria dreamed about the possibilities of a magnetoresponsive fluorescent...of improved response. Then, just when Andrew was about to suggest she deprioritize the project, Maria’s...with a strong commitment to exploration and collaboration. Andrew is a physicist with a background in ...potential applications.  Andrew is a frequent collaborator and is happy to give advice and support for ...interested in working on these questions or collaborating, the York Lab is eager to hear from you. Find...

... sample, the cut site will be aligned to the most 5’ cut site.     Indel Distribution In the Indel Distribution...deletions, the ability to analyze and interpret data about these types of edits is critical to the CRISPR workflow...Distribution and Traces - provides particular details about the indel profile of the edited sample. You can ...sequence base calls from the .ab1 file are also shown above each trace. The horizontal black underlined region...us know how it works for you! Read more details about the ICE tool in our bioRxiv paper. The new ICE CRISPR...guiding CRISPR researchers, she's either day-dreaming about exploring sci-fi worlds or exploring nature in the...

... your analysis? Lazic SE. BMC Neurosci. Jan 14;11:5. (2010) PubMed PMID: 20074371. PubMed Central PMCID...optimization within your experiment. Read on to learn more about the various controls that can be used for plasmid-based...: Expression Level of Gene X The results shown above are from a single experiment wherein Plasmid A (...be compared. The Empty Vector Control (mentioned above) could also serve as an important negative control...experimental cells. For example, in the experiment above, Plasmid A contains an shRNA that targets human ... times and on different days. Using our example above, we would test different preparations of Plasmid...

...et. al. An efficient gene knock-in strategy using 5’-modified dsDNA donors with short homology arms. Nat...CRISPR Featured Topic Page Learn about Non-Homologous End Joining Learn about Base Editing Read 3 Tips to ... post on increasing HDR efficiency to read more about optimizing this distance. Homology arms and types...screening strategies for your experiment. Also learn about Addgene’s knock-in plasmids for reagents to help...

... PMID: 25494202. PubMed Centra PMCID: MC4420636.  5. Sanson, Kendall R., et al. "Up, down, and out: next...Read this blog post about CRISPR Kinome Libraries available at Addgene Learn about the advantages of pooled...large-scale functional screens Read this blog post about genome-wide screening using CRISPR Resources on...libraries for the mouse genome! Other Resources Read about CRISPRi and CRISPRa from the NIH E-newsletter ...

...biotechnology 22.7 (2004): 346-353. PubMed PMID: 15245907. 5. Maertens, Barbara, et al. "Gene optimization mechanisms...that lead to these preferences and what can we do about them? Read on to find out!   Why do organisms have...understood, but some possible reasons include: Metabolic pressures - it takes cellular resources to produce...amino acid I’d like to encode,” but, as described above, not every region of a protein should necessarily...Effects on transcription (remember, it’s not all about translation - e.g. codon choice may interrupt transcription...

...necessary. To save time and money, I like to miniprep 5 mL of my AAV transfer plasmid cultures and then screen...a workspace with plasmid, not that I would know about this… A no template control (NTC) should always ... variability. Check out this post to learn more about different ways to titer AAV. Do you have any AAV...at Fred Hutchinson Cancer Center for teaching me about AAV production and purification! References 1. Aurnhammer...

...myelocytomatosis viral oncogene (myc), simian virus 5-derived epitope (V5), the synthetic peptide FLAG, ...enable accurate labeling of neurons. As discussed above, weakly expressed proteins in particular are difficult...resembles images of the deity (see comparison image above). The church of the Flying Spaghetti Monster is ...tags. 4. High-resolutions microscopy As indicated above, smFPs can be used with conventional microscopes...

...surprising-rules-you-should-know-regarding-linkedin-etiquette/5/#4050d70b1e9c https://www.linkedin.com/pulse/20140417174121...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...

...in seed plants." Plant cell reports 9.1 (1990): 1-5. PubMed PMID: 24226366. Lee, Min Woo, and Yinong Yang...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...

...linked to some diseases. It has been reported that 5-10% of human genes contain a region where a synonymous...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...

...or a magnet. 4) Unbound proteins are washed away. 5) The target protein is eluted.     In the first...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...

...Bimolecular fluorescence complementation (BiFC): A 5-year update and future perspectives. Biotech 53. https... 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...

...in MIC trays. Journal of Clinical Microbiology, 23(5), 959–961. https://doi.org/10.1128/jcm.23.5.959-961.1986...therefore prevent cell death. One of the cool things about amp is that it's exclusively toxic to dividing cells...antibiotic selection. If you feel like multiple of the above antibiotics would work for you, they likely do! ...

...biology 11.12 (2015): 917-923. PubMed PMID: 26575238. 5. Crivat, Georgeta, and Justin W. Taraska. "Imaging... 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 ...

...diseases. The researchers injected Cre-expressing AAV-5 into the DRN of a transgenic mouse harbouring a Cre-dependent...distant regions. They also don’t give any clues about the types of neurons involved or whether their involvement... Viral Vectors Resources on Addgene.org Learn about Addgene's viral service Find viral protocols and...

...biology 5.2 (2009): 118-126. PubMed PMID: 19136976. 5. Miyatsuka, Takeshi, Zhongmei Li, and Michael S. German...Additional Resources on the Addgene Blog Learn More About Fluorescent Protein Aggregation Catch Up On Your...