...microscopy with sequential multiple-view reconstruction (5). In order to image large tissue such as cleared rodent... shadows appear as stripes in the acquired image (5).  A way to get around this problem is to illuminate...protein (GFP) (6).   The Dodt and Frank Bradke laboratories later developed the 3DISCO (3-dimensional imaging...

...Edelhoch method. Int J Biol Macromol. 2015;77:260-5. doi: 10.1016/j.ijbiomac.2015.03.027. Epub 2015 Mar... in Colloid & Interface Science, Volume 19, Issue 5, 2014, Pages 438-449, ISSN 1359-0294, https://doi....validated, it’s ready for use! You can learn more about common antibody applications in our Antibodies 101...

...PAM sequence for recognition by QCascade; although 5’-CC-3’ is one of the best PAMs for VchINT, many other...by QCascade (Fig. 1B). The process also creates a 5-bp target-site duplication (TSD), a hallmark characteristic..., 2020), to multi-copy genomic integration of metabolic cassettes (Zhang et al. 2020); however, the technology...

... for Grant Writing 5 Reasons to Use Reddit for Science Communication Changing Labor Laws Bring Increased...record readership with over 75,000 views. Changing Labor Laws Bring Increased Postdoc Wages Our most popular... for anyone considering a postdoc. In “Changing Labor Laws Bring Increased Postdoc Wages,” Future of Research...thorough yet concise rundown of how recently enacted labor laws will likely increase postdoc earnings in the...changes quickly and harmlessly. Learn much more about the laws and their possible repercussions in Gary...This post leads readers to our latest podcast all about graduate student life and how establishing a thriving... labor laws...

...e-commerce company and establishing credibility Chapter 5 - Realizing that Addgene would be successful and the...repository’s future. If you’ve ever been curious about what it takes to start your own nonprofit, this ...Harvard University, where she studied diabetes and metabolism. Melina has worked at Genetics Institute in Cambridge...

... facilitate the isolation of modified cells; and (5) is compatible with the introduction of unique molecular...faithfully model the complex spatial, cellular, metabolic, and cytokine microenvironment that exists in ...LaFleur is a Postdoctoral Fellow in Arlene Sharpe’s laboratory at Harvard Medical School and is interested in...

...Retrograde (AAVrg), and volumes of 1 mL, 2.5 mL, or 5 mL. Please note that all orders will be shipped in...your browsing and requesting. If you’re curious about the service, read on.  Expanding our viral vector...

...DREADD Gether 83896 pAAV-hDlx-GiDREADD-dTomato-Fishell-5 hDlx Inhibitor DREADD Fishell 83897 pAAV-hDlx-GqDREADD-dTomato-Fishell...serotype from the Alla Karpova and David Schaffer laboratories. These viral vector preparations were produced...Janelia Research Campus viral project team in collaboration with David Schaffer (University of California...

...whose plans are unknown (up to 33% from 22%), a full 5% of postdocs who were expecting a salary increase ...Previous Article: Changing Labor Laws Bring Increased Postdoc Wages Read about the Advocating for Science... of $47,476 per year, under updates to the Fair Labor Standards Act (FLSA). This was due to the threshold...postdocs - and in some cases, faculty - have found out about what their institution is doing from Future of Research...Science Symposium and Workshop Learn about a Variety of Science Careers Resources on Addgene.org Find Career...

...reserved for questions. 5. Understand the job you are applying for It's not about you and how you will finally...members to discuss your research plans for the next 5-10 years. You will discuss the focus of your first... potential colleagues study. Think about how you might collaborate or, at the absolute minimum, can appreciate...the first sentence or two what your research is about and what you think you can accomplish. No matter...your seminar accessible. Let the experts ask you about the details and then you can display your deep knowledge...expert's question than to leave everyone confused about your research because you waded too deep into the...simple in principle The first question I usually get about chalk talks is, what is a Chalk Talk? It is a 60...

...place I could be happy with it. Cycle 5 - The Future! Figure 5: The mobile monitor and teleprompter... Production Specialist, I’m constantly thinking about ways to improve our productions to deliver the best...four months after I started with Addgene, I was about to have a check-in with my boss Jennifer. I had ... would be a couple months. Jennifer and I spent about an hour that morning scrambling. Everything we had...someone to be comfortable on camera while thinking about things like lighting, camera angle, and shot composition... allowing people to sit without having to worry about getting in close to mic them up. Monitor: I then...first video in our Intro to the Lab Bench series about PPE! We made this with a combination of in-lab filming...

...Elements of Acidithiobacillia. The CRISPR Journal, 4(5), 656–672. https://doi.org/10.1089/crispr.2021.0051...720–725. https://doi.org/10.1038/s41586-021-03886-5 Pinilla-Redondo, R., Mayo-Muñoz, D., Russel, J., Garrett...Cleavage during Type III CRISPR-Cas Immunity. Cell, 161(5), 1164–1174. https://doi.org/10.1016/j.cell.2015.04.027...Class 1 is the most abundant of the two, comprising about 90% of identified CRISPR-Cas in bacteria and nearly...CRISPR systems called CasPEDIA, with information about enzyme activity, experimental conditions, and more...

...creativity. Gallery Books. Boyes, A. (2019, November 13). 5 mental mistakes that kill your productivity. Harvard...Retrieved January 14, 2022, from https://hbr.org/2019/11/5-mental-mistakes-that-kill-your-productivity Finkelstein...brainstorming sessions.  Allow the group time to think about the problem before you meet. This holds the dual... meetings run smoother when people have thought about the issue ahead of time. Encourage dissent. The ... decisions while encouraging the group to think about alternative options. When a devil’s advocate is ...a few minutes every month dedicated to thinking about the future will improve your ability to innovate...to be able to continue our experiments. Thinking about these plans before you’ve run out of consumables...

...consider the following example gel. Figure 5: An example gel (left) with bands corresponding to...of western blots. Expert Review of Proteomics, 11(5), 549–560. https://doi.org/10.1586/14789450.2014.939635... this chain (blue) to infer or calculate things about earlier ones. In practice, multiple sources of error...condition 2.” By eye, the loading control appears about the same between the two conditions. But it isn’.... However, it throws both assumptions discussed above into question. Assumption #1: The loading control...searching the literature to see what is already known about your housekeeping protein in your context. If you...step from the POI blot, you don’t need to worry about the linear detection ranges of multiple targets....

... Protospacer Adjacent Motif (PAM sequence). The 3-5 nucleotide PAM sequence serves as a binding signal... Acidaminococcus sp. BV3L6 Cas12a, AsCpf1, uses a 5'-TTN PAM which makes it easier to target AT-rich genomes...Resources on the Addgene Blog  Read more about Cpf1 Read more about SaCas9 and AAV Check out our CRISPR Featured... and VRER) recognize novel PAM sequences (shown above). The VQR, EQR, and VRER Cas9 variants are capable...example, Cas9 from Staphylococcus aureus (SaCas9) is about 1 kilobase smaller than SpCas9, so it can be packaged...

...also takes into account that HDR is rare, usually 2-5% of the edited cells in our hands, these combined ...heterozygous event, the cut should ideally be around 5 to 20 bp away from the cut site. Lastly, if a heterozygous...mutations When thinking further about the distance relationship described above, we realized that, in addition...efficiency already drops by half and after only about 30 bp, it is no longer feasible to incorporate mutations...Additional Resources on the Addgene Blog Learn about High Fidelity Cas9 Variants How to Design Your gRNA...

...MgCl2, 4) increased concentration of dNTP, and/or 5) the addition of MnCl2. Disadvantages of this method...PMID: 25167058. PubMed Central PMCID: PMC4412028. 5. Araya, Carlos L., and Douglas M. Fowler. "Deep mutational... all desired codon changes (step 1 in the image above). A second universal primer is used to synthesize...first followed by the top strand. Both produced about the same number of positive clones. See Supplementary... of DNA degraded in the first digest (step 1 in above diagram). Remember that the BbvCI enzymes nick the...mutagenesis with the same protocol. What do you think about one pot saturation mutagenesis? How would you use...

...microbial cooperation, and antibiotic production. Chapter 5: The future of Synthetic Biology. Wide adoption of...synthetic yeast chromosome. Listen to learn more about Dr. Ellis, synthetic biology, synthetic chromosomes...Additional Resources from the Addgene Blog Learn about the Synthetic Biology Education Nonprofit BioBuilder...BioBuilder Plasmids 101: Golden Gate Cloning Learn About Working at a SynBio Startup in Our Interview with...

... mismatch off-targets were only toxic if they had 5+ off-target sites. Results from the screen also reproduced...Pubmed PMID: 23394947. Pubmed Central PMCID: 4299491. 5. Bassik, Michael C., et al. "A systematic mammalian...following unanswered questions about CRISPR screen design. Unanswered questions about genome-wide CRISPR/Cas9...Morgens et al drew some generalizable conclusions about off-target activity, demonstrate safe-targeting ...

...cells using compact expression cassettes. Sci Rep 5: . https://doi.org/10.1038/srep10777  Garneau JE, ...:97–110 . https://doi.org/10.1038/s41551-019-0501-5  Liu J-J, Orlova N, Oakes BL, Ma E, Spinner HB, Baney...Addgene Blog Learn more about Cas9 Variants and the Expanded CRISPR Toolbox Read about CRISPR enzyme Cpf1 ... and specific? The two AAV strategies described above showed successful target modification, indicating...