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We narrowed to 709 results for: abo.1

Showing: 341 - 360 of 709 results
  1. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

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    ...sequences are inserted into the genome. Figure 1: Self-excising cassette schematic from Dickinson et...):1028-34. doi: 10.1038/nmeth.2641. Epub 2013 Sep 1. PubMed. Find plasmids from this publication at ...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...
  2. Plasmid Cloning by PCR

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    ...has error rates that range from roughly 1 per 500bp to roughly 1 per 10 million bp depending on the polymerase...In its simplest form, PCR based cloning is about making a copy of a piece of DNA and at the same time...Additional Resources on the Addgene Blog Learn About Other Cloning Techniques Catch Up on All Things ...
  3. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

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    ...pipetting, he can be seen daydreaming about sunny days in the UK.   References 1. Bressan RB, Dewari PS, Kalantzaki...the tag sequence flanked by homology arms (Figure 1). In the first step of optimization, we started off...form the ribonucleoprotein complex (csRNP, Figure 1). We use in-house purified Cas9 protein, which, in...standard immunocytochemistry (ICC) assays.  Figure 1: Three ingredients of epitope-tagging experiments....ChIP-seq methods provide a wealth of information about how protein networks work in cells. A major bottleneck...two-part cr/tracrRNAs for CRISPR experiments (in collaboration with Integrated DNA Technologies, USA). These...commercial suppliers without investing time and labour on reagent preparation in the lab. Figure 3: csRNP-assisted...
  4. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

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    ...principles of fluorescent dyes/ligands Figure 1: Example structures of fluorophores, fluorescent dyes... dopamine D2/D3 receptors. Scientific Reports, 10(1), 21842. https://doi.org/10.1038/s41598-020-78827-...for bacterial imaging . Microbiology Spectrum, 12(1), e0369023. https://doi.org/10.1128/spectrum.03690...Biochimica et Biophysica Acta - Biomembranes, 1838(1), 15–33. https://doi.org/10.1016/j.bbamem.2013.09.005... A., Purkayastha, N., Flad, K., Schmidt, M. F., Tabor, A., Gmeiner, P., Hübner, H., & Weikert, D. (2020...
  5. Five Popular Model Organisms, Part 2

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    ...microbial research like metabolism. Transport, biofilms, and much more. Because of it’s BSL-1 status, E. coli ...oxygen. Additionally, its status as a Biosafety Level 1 microbe makes it a perfect candidate for introductory...with superior performance. Nature Communications, 9(1). https://doi.org/10.1038/s41467-018-04899-x Yokoyama... more about model organisms in the first Five Popular Model Organisms blog post Read more about plasmids...characteristics that make them ideal for studying in a laboratory environment. We’ve covered five model organisms...have been specifically developed to thrive in laboratory settings. One of its most attractive features...minutes, they are perfect for molecular cloning and metabolic engineering.  The E. coli genome has an extensive...
  6. QC Sequencing Technologies at Addgene

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    ...and recorded as a DNA strand is synthesized (Figure 1). Sanger sequencing requires four separate reactions...trace” file to determine the sequence. Figure 1: Overview of Sanger sequencing technology. Created...Technologies. Current Protocols in Molecular Biology, 122(1). https://doi.org/10.1002/cpmb.59  Wang, Y., Zhao,...sequencing was considered the gold standard until about 2008 when a new, higher-throughput sequencing-by-synthesis...
  7. Building and Validating Recombinant Antibodies: A Case Study

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    ...restriction digest and colony PCR.   Fig. 1: (A) Schematic of cloning, expression and validation... doi: 10.1371/journal.pone.0038313. Epub 2012 Jun 1. PMID: 22675541; PMCID: PMC3365890.... called NABOR (Neuroscience AntiBody Open Resource). The antibodies distributed through NABOR are recombinant...troubleshooting methodologies drew me to learn more about the process that was used to create this important...secondary antibodies are commonly used in research laboratories to detect the different subclasses, but there...
  8. Viral Vectors 101: The AAV Retrograde Serotype

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    ... There are two methods of neuronal tracing (Fig. 1): one to visualize neurons that send information to...significantly updated in 2023 by Jason Nasse.  References 1. Tervo, D. Gowanlock R., et al. "A designer AAV variant... Get Started with AAV Learn more about Lentivirus Find out More about Addgene's Viral Service Resources...We often think about the brain’s function in terms of its regions. But equally important is the way the...head bone’s connected to the neck bone,” but with about a hundred billion neurons and six hundred trillion...
  9. Predicting Adverse Reactions to Monoclonal Antibody Drugs

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    ...immunogenicity score (Mattei et.al., 2022).    Figure 1: Generation of immunogenicity scores by predictive...antibodies for the treatment of diseases. J Biomed Sci 27, 1 (2020). https://doi.org/10.1186/s12929-019-0592-z  ...determining the T-cell epitopes that will likely bring about unwanted immunogenicity. The 3-D models can be applied...limitations. Using methods together can inform decisions about in vivo application and clinical testing. This would...to drugs: adverse reaction terminology. WHO Collaborating Centre for International Drug Monitoring, Uppsala...
  10. Targeted Mutagenesis with EvolvR

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    ...activity degrades the displaced strand.   Figure 1: EvolvR consists of an error-prone polymerase (PolI...mutation rate and extend the editing window of EvolvR: (1) two additional mutations to PolI3M generated PolI5M...blog Learn about other CRISPR methods for bacterial genome engineering   Learn more about lambda red ... genetic variants in bacteria. Read on to learn about the key components of EvolvR and its potential applications...
  11. CRISPR 101: Any Base Transversion Editors

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    ...a glossary at the end of this post! Figure 1: Base transitions are edits between the purine bases...uracil DNA glycosylase. Nature Communications, 15(1), 4897. https://doi.org/10.1038/s41467-024-49343-5...uracil-DNA glycosylase. Nature Communications, 15(1), 6397. https://doi.org/10.1038/s41467-024-50012-w...In our last post, we talked about the first base transversion editors: CGBEs, or C → G Base Editors. ...apyrimidinic sites and abbreviated as AP sites. A collaborative team from the David Liu and Dali Li labs reported... CRISPR 101: Cytosine Transversion Editors Read about four base editor reporters Resources on Addgene.org...
  12. Evolution of Lab Techniques

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    ...blotting, named for its inventor Edward M. Southern (1), can be used to detect DNA identity, size, and abundance...hours at low cost with restriction mapping (Figure 1). Though some labs may prefer to send DNA directly... serotonergic system and behavior.     References 1. Southern, Edwin Mellor. "Detection of specific sequences...abundant in a sample. Whereas Southern blotting is labor intensive and requires a large amount of high-quality...DNA technologies that are now commonplace in laboratories. As scientific technologies rapidly progress...PMC4411503. Resources on the Addgene Blog Learn More about Restriction Cloning Get Tips on Analyzing DNA Sequencing...
  13. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

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    ...of an industry H-1B need to be aware of the April 1 deadline for companies to submit an application on...and non-profit organizations do not have the April 1 H-1B queue process or quota like industry, these organizations... National Interest Waiver. The three prongs are: (1) The foreign national’s proposed work endeavor must...-1B, you need to prove 3 threshold requirements: (1) that you have international recognition for your ... data and the list goes on and on. As you think about your career (and your future in general), keep in...in this post benefit from the applicant thinking about the process early, keeping good records, and planning...writing, compliance, core facility management, collaboration management, grant administration, clinical operations...
  14. New Optogenetic Tools for Cytoskeleton and Membrane Control

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    ...overcomes many of those limitations (Figure 1). Figure 1: Opto-katanin induces fast, localized microtubule...GTPase Signaling Toolbox. ACS Synthetic Biology, 11(1), 515–521. https://doi.org/10.1021/acssynbio.1c00604... Check out our Optogenetics Guide to learn more about how these modules work, and read on to learn how...to the cell membrane isn’t powerful enough, what about altering the chemical composition of the membrane...
  15. Plasmids 101: Screening Strategies Used in Plasmid Cloning

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    ...screening blog post for more information.   Figure 1: Result of a blue-white screen. Image from Stefan ...Because Sanger sequencing can usually sequence just 1 kb of DNA, custom primers are especially helpful in...often relies on antibiotic resistance. But what about identifying clones that contain your desired insert... restriction digest. The key is to be strategic about your enzyme choices. You’ll want to pick enzymes... for your PCR.  Find the details and learn more about colony PCR in this blog post.   Figure 2: ...Sanger sequencing or the other techniques mentioned above. In molecular biology, cloning a gene into your ... and tips Resources on Addgene.org Learn more about Sanger sequencing Read the Molecular Biology Reference...
  16. Transferable Skills Guide: External Collaborations

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    ... others, and organizing collaborations, keep these jobs in mind! References 1. Sinche, Melanie, et al...guide: Collaborating with others outside your organization also known as external collaborations. Interestingly... and collaboration management. For example, you may already be part of a scientific collaboration - show...friendly, and helpful. Finally, don’t forget about collaborations and projects you can do with others after...lead! Read our blog post about fostering communication in remote collaborations! Stay in touch: Maintaining...notes that remind me about the current state of a partnership. Every collaboration and partnership is different...favored by research-intensive careers. External collaborations are becoming increasingly important as more...
  17. Gendered Innovations: Why Does Sex of the Cell Matter?

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    ...healthcare system was $35.2 billion (or $140 per $1 invested in the trial). The goal of gendered innovations...is the goal of Gendered Innovations. This project: 1) develops state-of-the-art methods of sex and gender.... Gendered Innovations is an international, collaborative project—funded by the European Commission, the...research.  In addition to the studies referenced above, another excellent resource for research design ... whole human population. To learn a little more about where differences in effects on the sexes come from...cost. Especially in animal research, they worry about needing more study subjects and control groups. ...
  18. The time and cost required to make a plasmid

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    ...plasmid, based on the assumptions outlined in Table 1.   Of course, this is for the simple insertion of...any restriction enzymes or vector purchases (Table 1). Restriction enzymes and vector purchases can raise...cloning before you can start your experiment? What about all the reagents you need to order? Sometimes, it...dive deep into generating the data you’re excited about. When we were closing in on the one million plasmids...million plasmids shared than not!), we think it’s about time we updated this post. How much time does ...choose the right cloning method Find blog posts about scientific sharing here Download the Plasmids 101...
  19. Plasmids 101: Inducible Promoters

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    ...Hsp70, the heat shock releases heat shock factor 1 (HSF-1), which subsequently binds to heat shock elements...light-regulated two component systems designed by the Tabor lab. Which inducible promoter is right for me? Here...
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