... is letting people know it’s happening. Aim for 3-4 weeks advance notice for the first meeting so that...think about. Choosing a topic and gauging interest The first thing to do if you’ve thought about starting... discussions. Beyond the excitement of learning about new cutting edge science, researchers of all varieties...help stay current on new and emerging work by collaboratively discussing recent publications in a specific...up. At Addgene, we wanted to think more closely about the wealth of high-throughput sequencing (HTS) data...the paper.  Before the first journal club, think about the meeting itself A journal club should aim to ...published science. They also serve as a way to learn about work outside of one's own field in a structured ...

...Cas9 and the PITCh- and locus-specific gRNAs Step 4: Select for puromycin resistant cells Step 5: PCR ...If you follow CRISPR research, you know all about using non-homologous end-joining (NHEJ) to make deletions...plasmids, GFP knock-in cell lines can be created in about a month and a half, without the need for complicated...more predictable than NHEJ. As seen in the figure above, short (5-25 bp) regions of homology flanking a ...Directed Repair and Non-homologous End Joining Read About Other Genome Engineering Tools like Sleeping Beauty... Additional Resources on Addgene.org Learn All about CRISPR on Our CRISPR Guide Pages Find CRISPR Plasmids...

...nothing is covering it. Turning on the UV light 3-4 times a week for up to half an hour can help keep ...of the hood while work is in progress.Learn more about cell culture contamination in this post from the... bath constantly at 37°C for this purpose.Learn about the effects of CO2 on sample pH Cell morphology:... Set up a routine to split cells Don’t be lazy about splitting cells, instead try to form a routine. ...secretion of lactic acid (a by-product of cellular metabolism that is toxic to the cells), and need to be split...

...plate. We generally freeze at least 2 ampules each of 4 clones with different levels of expression of our ...specific genes. Many years ago, people in the laboratory became frustrated with using glass cloning rings...the hood. A 48-well plate is also prepared with about 200 ul of culture medium containing serum in each...incubator, additional growth medium is added to about halfway up each well. Tips for using the agarose...with the firmer 2% agarose gel solution discussed above for picking. Many thanks to our guest blogger Iris...

...DNA-guided enzyme." DNA-scissors (blog). Blog website. 4. Hsu, Patrick D., Eric S. Lander, and Feng Zhang. ...technology. To dig more into what researchers felt about NgAgo, I conducted a survey and asked researchers...successful, and the second section asked basic questions about the NgAgo protocols used by respondents (transfection...27136078. 2. Burgess, Shawn., et al. "Questions about NgAgo." Proteins & Cell (2016). 3. "NgAgo: Genome-editing...PMC4343198. Resources on the Addgene Blog Learn More about NgAgo Enhance CRISPR Targeting Specificity with ...

...291–300. https://doi.org/10.1016/0092-8674(86)90746-4 Munro S, Pelham HRB (1987) A C-terminal signal prevents...Do you ever wonder about the origins of some of the common techniques or tools you use in the lab? Take...times of the myc tag.  Origins of protein tags For about as long as the disciplines have existed, cell and...examples of new versions of Myc1-9E10 mentioned above basically involve taking the Myc1-9E10 variable ...

...Next-generation Sequencing. Molecular Therapy - Nucleic Acids 4:e260 . https://doi.org/10.1038/mtna.2015.32 Wright...viral genome sequencing (VGS). Why should you care about viral quality control? AAV vectors are produced ... let us know if you have any specific questions about viral DNA NGS you’d like us to discuss. We are also...or at [email protected] to answer your questions about our quality control process! The images in this...

... of HIV genes required for virus production (3 or 4). Third-generation vectors contain gag, pol, and rev...Plasmids 101 Learn about genome-wide screening with CRISPR and Lentivirus Read about the ClonTracr Library... the proper cell line is used; several common laboratory cell lines including 293 were immortalized with...

... 2000 Sep;81(Pt 9):2147-53. Pubmed PMID 10950970. 4. Lafon M. Rabies virus receptors. J Neurovirol. 2005...connected to the neck. Shortly after, we learned about organs and the jobs they perform. This became foundational...part of the answer to many biological questions about the brain. To address this, molecular tools to map...

...excessive nuclease activity in tissues (Li et al., 2018). 4. Selection marker while engineering viruses Acr genes...Bondy-Denomy et al., 2018). For example, AcrIIA4 mentioned above indicates that it’s the fourth Acr protein discovered...Cas9 Variant for Your Next Experiment Read more about Optogenetic Control of CRISPR   Resources on Addgene.org...

...mutagenesis in a diverse array of human cells. Sci Rep 4: . https://doi.org/10.1038/srep05105 Stephens CJ, ...capacity. For instance, with a packaging capacity of about 6 kb, the 1st generation of AdVs have already a ... CRISPR/Cas9 research! To find more information about the adenoviral delivery of CRISPRS/Cas9 using the...

... 520.7547 (2015): 278-280. PubMed PMID: 25877186. 4. Prior, Ian A., Paul D. Lewis, and Carla Mattos. "... cancer are yielding promising results in the laboratory, and Addgene is accelerating this research by...within that pathway that are of interest to their laboratory. A single mouse click on a gene of interest will...

...Characterization of Long Non-Coding RNAs. Non-Coding RNA. 2021;7(4):79. doi:10.3390/ncrna7040079 Horlbeck MA, Liu SJ,... to devise an experiment that can tell you more about its function.  Resources and references Resources...

...that they will switch to other newer topics every 4-8 weeks. These topics are typically cutting edge; ...company to collaborate whenever it faces an important challenge. The types of projects mentioned above are just...and why you might enjoy doing it. I’ll also talk about the skills you’ll need to do well as a healthcare...often work with such teams to help make decisions about R&D investments in either new internal efforts, ..., or through acquisitions of or collaborations with other companies. Medical Affairs groups work on building...building awareness among physicians and scientists about clinical research, drugs, the diseases they can ...professionals running the marketing, commercialization, collaborative, and financial functions associated with the...

...available for conjugation to a specific label (see step 4 for more information). Purify the Nanobody With A ...Control Protein Activity Learn more about protein tags Read about E. coli strains for protein expression..., if not better, than antibodies for all of the above listed molecular techniques, but they can also be...protein purification methods. Read on to learn more about nanobodies and how their structure and function ...between a fluorescent tag and the target antigen to about 2 nm, which can lead to higher resolution for super-resolution...nanobodies. If you’re interested, you can learn more about how nanobodies are identified in this review. If...

...guy from the lab was such a master drywall hanger?  4. Utilize your office space too Balance point number...early employees spent this remaining time by elaborately pranking each other, dressing in costume, and...frequently ask the question, “What’s your favorite part about working at Addgene?” Most Addgenies reply with an... confident that their teammates and project collaborators have completed their work thoroughly. It’s kind...rose-colored-glasses-esque picture of how everyone is kind and collaborative and all we do is have fun all day long, whistling...megaphone, or your company’s intranet. Don’t forget about the power of simple word of mouth - be sure to bring...company.        Additional Resources Read other posts about life at Addgene!Read more of our careers posts! ...

...the National Grants Management Association (NGMA). 4. Develop a professional development strategy  Build...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...

... and worked as a technical support specialist for 4 years. Read on for insights from my time in technical...disadvantages of this overall rewarding role. Learn about Other Addgenies and Their Careers in Our Career ...general experimental guidance, and resolve complaints about products. As you might expect, different companies...order to enhance the OD signals and then complain about high background. In many instances, variations ...protocol. And customers may tell all sorts of details about the application, just not the ones that matter. ...communication, and is particularly excited to hear about novel developments in the fields of gene and cellular...Academic v. Non-Academic Career Decisions Learn about Science Career Options Explore Tips for Technical...

...Cancer 2.4 (2002): 315-319. PubMed PMID: 12001993. 4. Lucey, Brendan P., Walter A. Nelson-Rees, and Grover...Learn All about Mammalian Vectors Read up on CRISPR Delivery Systems for Mammalian Cells Learn about the Dangers...Misidentified or cross-contaminated cells have been known about since 1966 when Stanley Gartler reported that most...there is a loss of reputation that can impede collaborations, especially with industry. Today we also have...The LGC Group also provides standards and has laboratories to carry our independent quality control for...culture contamination." Archives of pathology & laboratory medicine 133.9 (2009): 1463-1467. PubMed PMID...

...relevant to avian magnetoreception. Nature Chemistry, 8(4), 384–391. https://doi.org/10.1038/nchem.2447.  Lee...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...