...Snapshot: Negotiations When Cultures Collide: Managing Successfully Across Cultures by Richard D. Lewis ... partners and how can Addgene work with them to ensure we create a win-win situation? Learning about and...Whether you are serving on academic committees as a tenured professor or signing contracts when working for...Salary negotiations will be covered separately in a future post.   Resources Maria recommends: Getting...

...pitted against CRISPR head-to-head.  Exciting features of NgAgo First, unlike Cas9, NgAgo does not require...incubation at 55 °C - a dangerous, non-physiological temperature for mammalian cells and one which diminishes ...for the trickiness of NgAgo is the inability of mature NgAgo protein to load an ssDNA guide. It is possible...form a complex with a 5’-P-ssDNA guide at room temperature. Until that time, those who have been lucky with...using the Natronobacterium gregoryi Argonaute." Nature biotechnology (2016). PubMed PMID: 27136078. 2....

...from a unique promoter, multicistronic plasmids ensure that any cells that are positive for your marker...translation from an internal region of the mRNA. In the figure above, you can see that the IRES element acts as...you to spend less time researching plasmid basic features and spend more time developing the clever experiments...sequence derived from poliovirus RNA. Pelletier et al (Nature. 1988 Jul 28;334(6180):320-5.) PubMed. 2. A segment... mediated by T cell antigen receptor-CD3 ITAMs ensures effective negative selection and prevents autoimmunity...

...look at structure, impediments, the representation of women on committees, and how the culture of the ... promoters and terminators, but then they aren’t sure how to convert their own coding sequence to the ...science based around those technologies in the near future. Science on Twitter Addgene: You are active on ...Twitter account recently inspired by an academic in literature who gave a talk to the women’s research network...

...proteins associated with the IRF-1 locus. Figure 1: Schematic of enCHIP. Image from Fujita and Fuji...affinity purification in situ of regulatory elements (CAPTURE) (Liu et al., 2017). This system utilizes dCas9...through proteomics and the chromosome conformation capture (3C) technique. In addition to 3xFLAG and biotin...FSHD), a disease for which there is currently no cure (Campbell et al., 2018). Find plasmids from the ...Central PMCID: PMC6286148. Liu, Xin, et al. "In situ capture of chromatin interactions by biotinylated dCas9...

...an antibiotic resistance gene is a mechanism to culture only bacteria which have retained a functional ...formation on agar plates, less stable at high temperatures and in acidic environments.   Carbenicillin   ...can be successfully used for plant and bacterial culture, but some types of bacteria, such as SHuffle cells...all-in-one antibiotic — it is usable in eukaryotic cell culture, yeast, bacteria, and even plants! The zeocin resistance...., & Brooks, G. F. (1986). Effect of storage temperature and pH on the stability of antimicrobial agents...

...you don’t have plasmid ID numbers, or you aren’t sure if any plasmids from a paper have been deposited...plasmid search and found the results confusing, I’m sure you’ll find Addgene’s newly launched search engine...marker used to grow plasmid containing bacterial cultures Bacterial Resistance: Use this to choose the antibiotic...Editing brings us to a page with a basic overview and figure, followed by a convenient table of plasmids specifically...resources on the Addgene blog Learn more about other features on Addgene's website ...

...applications. Read on to learn more about the current (and future) applications of lentiviral vectors! Lentiviral...popular for targeting and tracing cells in the brain (Figure 1). 2nd generation lentiviral packaging system ...to create the 2nd-generation lentiviral system (Figure 2). The HIV genes that do remain are very important... Dull et al. created the 3rd generation system (Figure 3), which differs from the 2nd generation in two...Gluckman E, Bushman F, Hacein-Bey-Abina S, Leboulch P. Nature. 2010 Sep 16;467(7313):318-22. PubMed PMID: 20844535...

...are required for replication and transcription (Figure 1). All viral genes are removed and replaced by...transcription, as it would for a full-length viral genome (Figure 2). While it is technically possible to encode ... luciferase. Firefly luciferase can readily be measured to quantify (indirectly) transcription performed...result in significantly reduced activity (4). To ensure that you end up with appropriate plasmid ratios... not essential for virus replication in tissue culture, and the human RSV leader region acts as a functional...

... grant readiness, identify grant opportunities, secure grant awards, and manage grant projects and funds...management, we track grant opportunity deadlines, and we ensure that grant reports are submitted and project evaluations...field of grants. It is no surprise that the U.S. Bureau of Labor and Statistics shows that employment of...program didn’t stay in one place within the city infrastructure. My career path changed when I was asked if...She and her team at J. Miller & Associates have secured more than $70 million dollars in grant funding....

...plasmids of these systems to Addgene in the near future. Addgene: Has your lab been working on any other...development in biology and hope to develop more in the future. Involvement in the Scientific Community Addgene...papers submitted to journals. In addition, it is my pleasure to interact with my colleagues in meetings of ...institutes in different places have very different cultures. In Basel, I felt that the time flows much slower.... I hope I can help change the situation in the future. Addgene: What advice do you give to post-docs ...

... neurological defects. Nature 550:398–401 . https://doi.org/10.1038/nature24058 Tillotson R, Bird A (2020...can find the reagents and resources they need to cure Rett Syndrome.  If you’re new to Rett Syndrome, ...walking. They may exhibit slow head growth, develop seizures, cardio-respiratory dysfunction, distinctive hand...gene therapies to clinical trials in 2021. Figure 1: Schematic of the human MECP2 locus and MECP2...

...candidates have seen an actual chalk talk. Their first exposure to a chalk talk is usually their own. This is ...in the remaining time. The typical chalk talk structure is: What is the big question or mystery that ...livers for transplants. Growth of new livers in culture could resolve this shortage." What is the knowledge...Perfect your story and delivery. Design simple figures and diagrams to illustrate your vision. Practice...beyond the whiteboard. You may wish to create a sculpture or print a 3D model to better illustrate your ...

...can direct people to the AMA in the future and link to it in future blog posts on these topics. Others ... new questions and topics to write about in the future. The AMA was also cataloged for permanent reference.... I have a paid account which gives me all the features I need to save, organize and easily find the resources...on the internet or receive via email. The cool features are too many to describe here, but I have over...

...such as the brain, transparent so that the 3D architecture of a fluorescent neuron is visible. Chemical... al. "Improved tools for the Brainbow toolbox." Nature methods 10.6 (2013): 540. PubMed PMID: 23817127...neural circuit analysis in Drosophila melanogaster." Nature methods 8.3 (2011): 260. PubMed PMID: 21297619.... arises from expansion and dedifferentiation of mature pigmented melanocytes." Cell stem cell 21.5 (2017...of fluorescent proteins in the nervous system." Nature 450.7166 (2007): 56. PubMed PMID: 17972876. Pan...

...formation requires a fully folded beta-barrel structure, followed by an intramolecular reaction that generates...shown) remains fluorescent in acidic organelles (Figure 1), showing that its acid tolerance is maintained...specific phosphoinositides (Varnai and Balla, 2008) (Figure 2). RNA production can be detected by using a MS2...specifically detects a specific RNA stem loop-structure (Querido and Chartrand, 2008). Finally, a nuclease-deficient...Quantitative assessment of fluorescent proteins." Nature methods 13.7 (2016): 557. PubMed PMID: 27240257...

...interactions found in nature. Once the bond is formed it is unaffected by changes in pH, temperature or organic ...Discovery of the biotin-avidin interaction   Figure 1: Biotin is enriched in egg yolk while its binding...affecting the avidin binding site. With all of these features it isn’t surprising that scientists began to exploit...with biotin ligase and monovalent streptavidin.” Nature Protocols 3.3 (2008): 534–545. PubMed PMID:18323822...

...both targeted alleles when gene editing.   Figure 1: Techniques to improve editing efficiency. ... RNAs wisely (and let them cut close).   Figure 2: Relationship between cut site to mutation distance... one or the other on each target site.   Figure 3: Optimal cut site to mutation distances for heterozygous... and heterozygous mutations using CRISPR/Cas9." Nature 533.7601 (2016): 125-125. PubMed PMID: 27120160...scarless genome editing in stem cells using CORRECT." Nature protocols 12.2 (2017): 329-354. PubMed PMID: 28102837...

...standardized leader peptide prevents secondary structures in the mRNA encoding your gene of interest from...in-vitro assays: Koch, Daniel, et al. "A pull-down procedure for the identification of unknown GEFs for small...monitoring sequence dynamics under various selective pressures: Miller, Corwin, et al. "Adaptation of Enterococcus...applications of PUF protein fusions reported in the literature include RNA detection, RNA cleavage, and translational...endonucleases with customized sequence specificities." Nature communications 3 (2012): 1147. Cooke, Amy, et...

...few hours at low cost with restriction mapping (Figure 1). Though some labs may prefer to send DNA directly...computer program) can then reproduce the DNA sequence (Figure 2). Accuracy and accessibility led to the widespread...reactions and may use Sanger sequencing. Looking to the future Southern blotting, restriction mapping and sequencing...sequence an entire genome for under $1,000 (10). The future of sequencing should bring efficient, low cost ...large amounts of DNA quickly and accurately. We're sure to see many improvements in a variety of biological...