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We narrowed to 397 results for: abo.4

Showing: 141 - 160 of 397 results

  1. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...PMID: 25531807. PubMed Central PMCID: PMC4273606. 4. Improved vectors and genome-wide libraries for CRISPR...Shalem O, Zhang F. Nat Methods. 2014 Aug;11(8):783-4. PubMed PMID: 25075903. PubMed Central PMCID: PMC4486245...gene therapy applications. Read on to learn more about the current (and future) applications of lentiviral...do both, Addgene depositor Didier Trono and collaborators turned to the lentivirus HIV-1, well-known to...Libraries Enable Genome-Scale, Knockout Screening Read about Genome-Wide Screening Using CRISPR/Cas9 Additional...Addgene.org Check out Our Lentivirus Guide Learn about Virus Biosafety Find Lentiviral Vectors for Your...

  2. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ...approach using a single crRNA array to express up to 4 crRNAs. Zetsche et al. also suggest that Cpf1 may ...Kleinstiver et al. saw no decrease in Cpf1 activity with 4-6 base deletions at the 3’ end of the target sequence...genome editing than Cas9 - read on to learn more about Cpf1 and check out our CRISPR guide for a refresher...mediated cleavage are 42-44 nucleotides in length, about the same size as Cas9’s, but with the direct repeat... Cpf1 was first identified, we didn't know much about its on-target and off-target editing efficiency....CRISPR system proved we had a lot more to learn about CRISPR biology. Later work, like the adaptation ...

  3. Negotiating Work and Life: How to Find the Joy

    Type
    Blog Post
    ...people have regular schedules. Work 8 to 5 or 7 to 4 or maybe 7 to 6 and 8 to 10 or whatever works for ...carefully and with attention to those around you #4 Thou shalt not bear the burden alone You, alone, are...on the Society for Laboratory Automation and Screening (SLAS) Electronic Laboratory Neighborhood.   Work...worry less about how they will handle an emergency and this will make you much more confident about their ...

  4. Plasmids 101: Codon usage bias

    Type
    Blog Post
    ...specific amino acids. This is possible because there are 4 bases (A, T, C, and G) and 3 positions in each codon...PubMed PMID: 2265611. PubMed Central PMCID: PMC552224. 4. Gustafsson, Claes, Sridhar Govindarajan, and Jeremy...that lead to these preferences and what can we do about them? Read on to find out!   Why do organisms have...understood, but some possible reasons include: Metabolic pressures - it takes cellular resources to produce...amino acid I’d like to encode,” but, as described above, not every region of a protein should necessarily...Effects on transcription (remember, it’s not all about translation - e.g. codon choice may interrupt transcription...

  5. Design Tips for Prime Editing

    Type
    Blog Post
    ...to indels (Anzalone et al., 2019). Figure 4: Editing the PAM prevents the prime editor from re-engaging... 149–157. https://doi.org/10.1038/s41586-019-1711-4 Chen, P. J., Hussmann, J. A., Yan, J., Knipping, F... primer binding site, starting with a length of about 13 nt (Anzalone et al., 2019). Primer binding sites...of reverse transcriptase template, starting with about 10–16 nucleotides. For longer templates, it is even.... Test multiple nick sites, starting with sites about 50 bp upstream and downstream from the prime editing...

  6. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...harvested and purified. In total, this process takes 4-7 days, not counting the time needed to expand the...completed in one long day, or the virus can be stored at 4 °C and buffer exchanged the next day. Refer to the...a workspace with plasmid, not that I would know about this… A no template control (NTC) should always ... variability. Check out this post to learn more about different ways to titer AAV. Do you have any AAV...at Fred Hutchinson Cancer Center for teaching me about AAV production and purification! References 1. Aurnhammer...

  7. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    ...designed to detect resulting in false positives [2, 4]. Solutions to the polymer problem There are some ..., 1999. 17(10): p. 969-73. PubMed PMID: 10504696. 4. Ohashi, T., et al., An experimental study of GFP-...sold by Clontech and is in a freezer box in just about every lab in the world, is not inert. In all seriousness....11 mM [2]. Following the simple logic outlined above, you might think EGFP is unlikely to form dimers...affinity assays used in the papers describing the FPs above including molecular sizing columns and native gels...

  8. Tips for arabidopsis transformation

    Type
    Blog Post
    ...plant synthetic biology’, Frontiers in Plant Science, 4. doi: 10.3389/fpls.2013.00339. PubMed PMID: 24032037...PMID: 25902487. PubMed Central PMCID: PMC4426443. 4. Nakagawa, T. et al. (2007) ‘Improved Gateway Binary... J. (2002) Arabidopsis : a laboratory manual. Cold Spring Harbor Laboratory Press. Available at: https...https://books.google.at/books/about/Arabidopsis.html?id=IfZAMNPWVk4C&redir_esc=y (Accessed: 21 September ...

  9. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    ...fusions to 3 or more HA tags. 4. High-resolutions microscopy As indicated above, smFPs can be used with conventional...2009.12 (2009): pdb-top63. PubMed PMID: 20150100. 4. Shaner, Nathan C., George H. Patterson, and Michael...enable accurate labeling of neurons. As discussed above, weakly expressed proteins in particular are difficult...resembles images of the deity (see comparison image above). The church of the Flying Spaghetti Monster is ...

  10. An Interview with Connie Cepko - Gene Therapy, Plasmid Tools and Insights to Success

    Type
    Blog Post
    ...scaffolds for cell-specific gene manipulation.” Cell 154(4) (2013): 928-939. PubMed PMID: 23953120. PubMed Central... Cepko. Listen to the podcast below to hear all about the research being conducted in Professor Cepko'...and mentoring styles. If you'd like to learn more about Professor Cepko's recent work on developing GFP-...the time to talk to us and look forward to hear about more exciting discoveries from her lab. Special ...

  11. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    ...cohesive end ligation by this enzyme for all 3- and 4-base overhang sequences under standard reaction conditions...identification of mismatch-prone pairings. Using these 4 bp overhang observations, accurate “high-fidelity”...Inserts, either precloned or in amplicon form (shown above) are flanked by BsaI sites in an orientation that...Check out NEB's work on ligase efficiency Learn about other plasmid cloning techniques Resources on Addgene.org...

  12. The Developmental Studies Hybridoma Bank: Over 25 Years of Antibody Sharing

    Type
    Blog Post
    ...exhibit increased avidity to high profile antigens; and 4) generating mAbs against new cancer biomarkers. We...antibodies to be used as a powerful reagent in the laboratory.   The DSHB focuses on monoclonal antibodies,...Hybridoma Bank (DSHB) since 1996.       More Blog Posts about Scientific Sharing: The 10 Most Distributed Plasmid...

  13. Hot Plasmids: Winter 2025

    Type
    Blog Post
    ...lentiviral packaging as a single payload (Figure 4). Figure 4: ONE-GO biosensors measure the formation...synaptic events By Brian O’Neill The Lin Lab and collaborators recently created ASAP5, a faster and more responsive...

  14. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    ...Thousands of Reporters Integrated in Parallel. Cell, 154(4), 914–927. https://doi.org/10.1016/J.CELL.2013.07.018... 149–157. https://doi.org/10.1038/s41586-019-1711-4 Chen, P. J., & Liu, D. R. (2023). Prime editing for... al., 2023). We trained our models in close collaboration with the Krauthammer lab (University of Zurich...

  15. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    ...:3499-3510. doi: 10.1002/bit.27776. Epub 2021 May 4. PMID: 33811659. Pace CN, Vajdos F, Fee L, Grimsley...absorption coefficient of a protein. Protein Sci. 1995 Nov;4(11):2411-23. doi: 10.1002/pro.5560041120. PMID: 8563639...validated, it’s ready for use! You can learn more about common antibody applications in our Antibodies 101...

  16. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ...buffer. Additionally, protein lysis should be done at 4 °C or on ice, whichever is more practical. If your...protein-based blockers degrade fairly quickly, even at 4 °C, so when in doubt, make up a fresh batch.  Antibodies...temperature and time. Though the most common conditions are 4 °C overnight or room temperature for 1–2 hours, there...protein/sample loaded Incubate primary antibody at 4 °C overnight Incomplete washing Ensuring wash...re in the right place! In this post, we’ll talk about western blot optimization and troubleshooting.   ...can be informative, so spend some time thinking about which type of gradient can help answer your particular...overexpression. Before starting a western blot, think about how you’ll interpret the data and what level of ...

  17. Podcast: Interview with Addgene's Founders

    Type
    Blog Post
    ...tech transfer and the adoption of the UBMTA Chapter 4 - Developing Addgene as a modern e-commerce company...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...

  18. Make a Splash: Notions of Scientific Impact Are Evolving

    Type
    Blog Post
    ...of 39 Scientific Impact Measures" (2009) PLOS One 4(6): e6022. doi: 10.1371/journal.pone.0006022 Aragon...Of course, all of you toiling away in laboratories this holiday season want the work you are doing to... has to do with online accessibility. It is now about as easy to find a paper in a lesser-known journal...

  19. 2018 #DeckTheLab Contest Winners

    Type
    Blog Post
    ...in labcoats (@ladiesinlabcoats) on Dec 19, 2018 at 4:36am PST Buddy the Elf from Matthew Sikora’s lab...more (stay tuned for a few blog posts coming up about sustainability in the lab!). The @ladiesinlabcoats...

  20. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    ...the sequence isn’t in place yet.     Figure 4: S. pyogenes Cas9 in the apo state (PDB ID 4CMP, top...641–656. https://doi.org/10.1038/s41579-022-00739-4 References Bravo, J. P. K., Liu, M.-S., Hibshman, ...sequence. Nature Structural & Molecular Biology, 29(4), 395–402. https://doi.org/10.1038/s41594-022-00756...nascent heteroduplex and abolish cleavage (Pattanayak et al., 2013). As mentioned above, however, Cas9 sometimes..., 2022), indicating that there is more to learn about this process!      Figure 3: Crystal structure...rest of Cas9 and is highly flexible; it can rotate about 140 degrees between inactive and active conformations...conformational changes that make Cas9 such a valuable laboratory tool are still an area of active research. New...
Showing: 141 - 160 of 397 results