...be collected from heterozygous individuals. Only 1/4 of the descendants of the heterozygous plants, therefore... is difficult for new members of the lab or collaborators unfamiliar with these mutant plants. Using RUBY...resistant disease. (RUBY-red: You can’t beet it!) About the Author  Andrew Willoughby is a graduate student...

...so it might lend itself more readily to a switch. 4. Do you want more autonomy?  Before my master’s, I... Resources Resources on the Addgene blog Learn about career planning resources What Do I Do Now? Academic...

...for individual cells. Each cell is labeled with 3-4 tags to increase the diversity of the CellTag library...Additional resources on the Addgene blog Learn more about induced pluripotent stems cells (iPSCS) and reprogramming...

...2018. PMID: 29735997 bioRxiv preprint posted July 4, 2017 A new method to identify proteins occupying... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here. Listen...

...within a few hundred nanometers of the coverslip (4). Essentially, an optical section is achieved that...biophysics 48, 178-243 (2015). PubMed PMID: 25998828. 4. Axelrod D. Total internal reflection fluorescence...call Epi-fluorescence microscopes. (See left image above for an example image). Point Scanning Confocal Microscopy... plane to reach the detector and all light from above or below is excluded. Parallelized Confocal Microscopy...due to limitations in light penetration mentioned above (5). Thick static samples Ex: Fixed tissue sections...sectioning. Point scanning confocal microscopes (see above) often provide the highest quality images for this...

...Cre-mediated marker excision, it takes approximately 3-4 weeks to obtain a homozygous, outcrossed knock-in ...another similar co-conversion marker (repair of a zen-4(cle10ts) allele), increasing experimental options....learn more about genome editing with CRISPRs? Check-out Addgene's CRISPR Resources Learn about gRNA design...interest, I encountered low efficiency (0.13%) and laborious handling (screening 768 F1 animals). Recovering...

...become your article title.       Figure 4: Add plasmids to the article by clicking “Add Plasmid... cloning information for the gene or insert. Step 4: Growth and Distribution Please indicate which strain...annotate on this page. Figure 7: Enter information about the gene/insert(s). Step 3: Cloning Information ...

...grant writing Job changes can happen regularly (every 4-6 years is common), but this can also bring new opportunities—projects...students Collaborations may be less “complicated” - for example, easy to initiate a collaboration without...up the pace Constant worry about grants and renewals What is different about non-academia? Scientific...make a decision about something like this without sufficient data? Starting to learn about the diverse options...with scientists just like you What is different about academia? Publishing in peer reviewed journals ...without having to worry about proprietary information Work hours are long, but there is more flexibility with...paths to pursue. Spend some time really thinking about your motivations and what parts of being a scientist...

...your role as a lab leader, and then on to Chapter 4 for advice on staffing your lab. Let us know if you...start their own academic research laboratory. How does one even go about starting a lab? Obviously, funding...always helps. But how does one build a strong collaborative research team motivated by similar visions and...booklet. What has been the most surprising thing about becoming a PI? Tom Ellis has a one word answer: ... has to deal with. My inbox currently stands at about 65,000 messages and dealing with email can be half...the first couple of hires to build a strong, collaborative team. And take advantage of the many resources...Medical School) for taking the time to speak with us about adding new scientists to their labs. Further Reading...

......perhaps you get the idea. What will you start? 4. Do informational interviews (on a schedule) I hope...mentoring group (see above). However, now that we are all so good with video calls, how about trying your hand...blog or podcast, initiated and maintained a collaborative project, served on the board of a professional...appointments with people all over the world. Think about contacting alumni of your lab or university or use...suggest you speak with. See this post that’s all about informational interviews, for more details on how...progress on both building relationships and thinking about future career options. 5. Write a grant - even if...

...Schmidt H, Krause MW, Seydoux G. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014...just one injection step! The entire protocol takes about 2-3 weeks but requires less than eight hours worth... this species, namely, the robust HDR mentioned above, an accessible germline, and multiple selectable...modular system that requires very little hands-on labor and could facilitate the tagging of every protein...

...Next-generation Sequencing. Molecular Therapy - Nucleic Acids 4:e260 . https://doi.org/10.1038/mtna.2015.32 Maynard...share the procedures and Python scripts with laboratories and viral vector cores routinely producing AAV...on Addgene.org Find AAVs at Addgene Learn more about Addgene's viral service Find protocols for working...

...same name. The tag has a net negative charge with 4 negatively charged and 1 positively charged amino ...Tags have a lot of major benefits – as discussed above – but there are a few instances when they should... hindrances, and destabilization can be brought about by the tag so it’s important to validate tagged ...

...emission maximum is 675 nm. mCarmine is monomeric, 4-5 fold brighter than mNeptune684, and has a favorable... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.   ...emission maxima toward the red end of the spectrum (above 600 nm) and increasing the brightness. Arne Fabritius...

...formation as well as increased Zfp462 expression within 4 hours of blue light induction (6-fold faster than ... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.    ...neurons. Earlier this year, the Hegemann lab and collaborators report their discovery of a new family of seven...

...the Syntaxin3 gene was the winner. We used the next 4-5 weeks to work on cloning the Syntaxin3 gene in a...intern coordinator shortly after hearing her talk about Addgene’s internship opportunity taking place next...

...Fact 19. https://doi.org/10.1186/s12934-020-01321-4   Kolodner R (1980) Genetic recombination of bacterial...molecular biology and viral vector protocols Learn about the common lab E. coli strains Find tips for viral...

... OR you can store the reaction at -20C overnight. 4. Transform TOPO Cloning Reaction into Competent Cells...in mind that Taq polymerase has an error rate of about 1 in 3,500 bases. Typically polymerases with proofreading...

..., Johansson M. Nat Biotechnol. 2001 Aug;19(8):713-4. PubMed.  ... a protocol has been established) may be less laborious than viral purification, and it does not raise...

...Translation of Single mRNA Molecules In Vivo.” Cell 165(4) (2016): 976-89. PMID: 27153498 PMCID: PMC4889334 ...Additional Resources the Addgene Blog Learn More about SunTag Catch up on Your GFP Background Choose the...