...scoop from Addgenie A. Max Juchheim in this post. 3. Plasmids 101: E. coli Strains for Protein Production...eyes open for any new technologies that will bring about the next revolution in biological research. We’re...Plasmids 101: Control Plasmids Need help thinking about the types of controls you’ll need for your next ...Addgene Browse Our CRISPR Resources Learn More about Viral Vectors Browse the Repository  ...

...International Publishing. https://doi.org/10.1007/978-3-319-07758-1_2 More resources on the Addgene blog Viral... using biorender.com   If you’re new to talking about viruses, here’s a quick tip on terminology. “Virus...have a growing number of Viral Vectors 101 posts about each type, so be sure to check our blog, or our ...

...fluorophore's excitation spectra around 640 nm (Figure 3), we see that APC-Cy7 is excited at a lower efficiency...cannot be excited at this wavelength. Figure 3: Excitation and emission spectra for APC and APC-Cy7...spectrum is located in a range of wavelengths of about 380–700 nm. Photons of lower wavelengths are higher... 650 nm, APC-Cy7's global excitation maximum is about 750 nm. If we were to use a laser that excites at...optimal excitation happens at higher wavelengths of about 750 nm. Source: https://www.bdbiosciences.com/en-us... we start building your first panel, we'll talk about one last principle of color emission. As you can...our blog post on color compensation to learn more about them. Alright, let's build your first panel! Designing...

...three systems to analyze single cells from a mixture (3): An optics system A fluidics system An electronics...expression. A routine flow cytometry experiment uses 2 or 3 different fluorescent colors each measuring a different...biosafety cabinet with cloning rings or following labor-intensive limiting dilution protocols? If this sounds...fluorescent microscopy must be used. To learn more about the most suitable fluorescent microscopy technique...a laser and the instrument collects information about the cell’s size, complexity and fluorescence intensity...captured by detectors and provides useful information about the cell. Light that continues in the same direction... forward scatter (FSC) and provides information about the relative size of the cell; larger cells produce...

...preparation, and lead your own research project.  3. Do you prefer being a generalist? Working on a variety... Resources Resources on the Addgene blog Learn about career planning resources What Do I Do Now? Academic...

...Palindromic Repeats, in Pubmed returned 728 articles (3/12/2014). With so many options for CRISPR plasmid ... gRNA design tools Need more questions answered about CRISPRs? Check out the full list of 16 FAQs answered...

...identifier 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...

...tagged with different fluorescent proteins or dyes. 3. Two-photon microscopy Two-photon excitation microscopy...PMID: 18718768. PubMed Central PMCID: PMC2569002. 3. Egen, Jackson G., et al. "Macrophage and T cell dynamics...fluorescence microscope where the light source is mounted above (epi) the specimen and the excitation light passes...technique that allows imaging of living tissue up to about one millimeter in depth. Two low-energy photons ...response, but the study of these dynamics was still laborious as epifluorescence microscopy wasn’t able to give...skin. Two-photon intravital microscopy. The video above depicts the infiltration of neutrophils expressing...

...(i.e. optical sectioning (2) or super-resolution (3). Fluorescence microscopy techniques   Fluorescent...methods 2, 920-931 (2005). PubMed PMID: 16299477. 3. Eggeling C, Willig KI, Sahl SJ, Hell SW. Lens-based...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...

...-mediated marker excision, it takes approximately 3-4 weeks to obtain a homozygous, outcrossed knock-in...produced a higher editing efficiency, while for edits 3’ to the PAM sequences corresponding to the opposite...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...

...one company, likely will need to move around every 3-6 years—a short and exciting ride Complicated matrix...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...

...and New Faculty" downloadable book. Skip to Chapter 3 for tips on how to develop your role as a lab leader...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...

...ready for the live audiences in your future.     3. Start something In almost every career talk I present...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...

...and (3) development of tools for data analysis/visualization using R/Shiny. She is passionate about open...section-specific comments, I re-read what I wrote about the paper after my initial read, adjust my language...wish for The last thing that I wanted to mention about the peer-review, is that as a peer-reviewer, you...each peer-review, I try to mention what I liked about this paper and recognize the hard work that the ...review article Resources on Addgene.org Learn about depositing plasmids pre-publication Find molecular...

... of the Brunello library are also shown in Figure 3. I hope you’ll agree with me that they look very similar... and we thought it would be a good time to talk about the amplification and verification processes we ... DNA. These reactions should allow us to obtain about 100 to 1,000 bacterial colonies per individual plasmid...from the count_spacers.py file from Feng Zhang’s laboratory. There are a number of other analysis scripts...you have any tips, input, or stories of interest about library use, let us know in the comments! Additional...NGS Sequencing to Validate AAV Preps Learn More about Pooled CRISPR Libraries Resources on Addgene.org...

...Brandon Sanderson "The Mistborn series is the first 3 books of a larger epic of fantasy novels. Set in a...resources on the Addgene blog Learn about company culture at Addgene Read about other lunch activities at Addgene...scientists bring biases to research and don't collaborate with others to bring diverse perspectives. Everyone...Science is a terrifying and equally good wake up call about the science of race."     - Joanne Kamens, Executive...easy reads, some are hard. Some I’ll never think about again, some I loved so much I’ll force them on everyone...

...page. Figure 7: Enter information about the gene/insert(s). Step 3: Cloning Information Please enter ... of 10 or more plasmids.       Figure 3: If submitting plasmid(s) online (left), choose to...

...et al., 2014) (Addgene #104106 - #104108) (Figure 3) that can be introduced using in utero electroporation...Additional resources on the Addgene blog Read about Brainbow Learn about using AAVs for neuronal tracing Visit...page Additional resources on Addgene.org Learn about Addgene's viral service See our fluorescent protein...

...stressed or aging cell populations. Figure 3: Micrographs of wild type (top panels) and cells with... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.  ...non-overlapping cells. Until now, tools for doing this were labor-intensive and required using manipulations such ...

...thousands of loops. These loops not only provide a 3-D structure for packing genomic DNA into nuclei, but... 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...