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Showing: 511 - 540 of 846 results

  1. To Each HIS Own

    Type
    Blog Post
    Published
    March 2, 2023, 2:15 p.m.
    ...Much of today's biological research requires a close examination of specific proteins within a system...without eluting the target protein and needs to be empirically determined. In addition, some metal ion substrates...function. B, The string of histidines in the tag is typically between 6 to 9 residues long. The small size of...Enfors SO, Persson I, Nilsson B and Uhlen M. Large-scale affinity purification of human insulin-like growth...

  2. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    Published
    Feb. 16, 2023, 7:05 p.m.
    ...protein expression in mammalian cells: easy to use biological tools are abundant! In this post, we will review...that! CRISPR interference (CRISPRi) targets a catalytically dead Cas9 fused to a transcriptional repressor... protein, then these systems need to be site-specifically knocked in. This can be labor intensive and ...and target-specific protein degradation. Nature Chemical Biology. May 2018. 14(5): 431-441. PubMed PMID...

  3. Summer SciComm: Preprints

    Type
    Blog Post
    Published
    Aug. 16, 2022, 1:15 p.m.
    ...arXiv.org), but have been only adopted by biological and biomedical scientists (see: bioRxiv.org or medRxiv.org...Preprints can be read freely by anyone and are typically free of charge for authors to post. They can be... be assessed and commented on by readers, typically peers in the field, and updated by the authors over...

  4. Antibodies 101: Selecting the Right Antibody

    Type
    Blog Post
    Published
    March 15, 2022, 1:15 p.m.
    ...exciting new project aimed at understanding how a critical cellular pathway is regulated. You’ve read all...do you know about the vendor and the vendor’s technical support? An antibody from a vendor that you use...frequently, with a responsive and knowledgeable technical support team, is a safer bet than one from a large... all validation is equal and you should think critically about what data is and is not shown on an antibody...

  5. RNA Extraction Without A Kit

    Type
    Blog Post
    Published
    March 24, 2020, 1:15 p.m.
    ...widely known of these products is TRIzol® (also called TRI Reagent®, RNAzol®, QIAzol®  and sold by many... within cells or tissues- they don’t actually chemically stabilize the RNA molecules (Allewell and Sarma... MacDonald RJ, Rutter WJ (1979) Isolation of biologically active ribonucleic acid from sources enriched...Guanidinium Thiocyanate–Phenol–Chloroform Extraction. Analytical Biochemistry 162:156–159. https://doi.org/10.1016...

  6. Reproducibility for Everyone: Lessons from an Open Science Collaboration

    Type
    Blog Post
    Published
    Feb. 28, 2019, 1:34 p.m.
    ... helped by expanding this team, leading monthly calls, and planning next steps. April Clyburne-Sherin ...first iteration of these workshops took place at Caltech in Jan 2018 and involved presentations and a handout...Not everyone in the group was able to make it physically to instruct the workshop at the conference, but...have the materials and resources to run their own local workshops was a major goal for the team last year...

  7. Oh, The Places You Can Go: Careers in Science Communication - Writing for a Research Institute

    Type
    Blog Post
    Published
    Feb. 2, 2018, 3:20 p.m.
    ...because her skill set wasn’t matching up with the local job postings. “There were jobs that I didn’t seem...team meetings and Susan is assigned to cover the Clinical Research Division at Fred Hutch, so she attends...work include: “Listening. Evaluating evidence. Critical thinking. Being able to learn things quickly. ...like she did for the Northwest Association of Biomedical Research. She does, however, recommend using ...

  8. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

    Type
    Blog Post
    Published
    Dec. 16, 2015, 3:30 p.m.
    ... Not only is CRISPR having effects on various biological disciplines, the base technology itself is constantly...at non-target sites. Means to decrease these so-called “off-target effects” have included using a Cas9... the Broad Institute, the Joung lab at Harvard Medical School, and the Doudna Lab at Berkeley set out ...afforded by gRNA binding at non-target sites would theoretically decrease off-target cutting. Figure 1...

  9. A Look at Addgene's QC Process

    Type
    Blog Post
    Published
    May 6, 2025, 1:15 p.m.
    ...plasmids. These are prone to error — both human and biological. Even the most high-fidelity polymerases can ...the correct inserts, tags, promoters, and other critical functional components. Fun fact! Some older plasmids...sequences. These sequences can also be confirmed or theoretical. No matter the level of information that is provided...the raw NGS reads and reassemble using a program called Geneious. Here, the raw reads are mapped against...

  10. Antibodies 101: Choosing the Right Isotype

    Type
    Blog Post
    Published
    April 6, 2023, 3 p.m.
    ...choosing antibodies   Complexity is a key factor Biological systems tend to be complex with a variety of ...multiple targets in parallel. In these cases, it is critical that users choose antibodies that are easily distinguishable...staining multiple targets indirectly, you must strategically plan primary/secondary antibody pairing such...Fisher, S. K. (1993). Chapter 15 Postembedding Immunocytochemical Techniques for Light and Electron Microscopy...

  11. Corralling and Cornering the Goldilocks Antibody

    Type
    Blog Post
    Published
    Aug. 9, 2022, 1:15 p.m.
    ...researchers to identify and isolate very specific biological targets. However, as popular and useful antibodies...antibodies, creating a potentially unlimited supply of identical antibodies (See Fig 1.). rAbs are a relatively...has specific binding and has a strong signal. We call the unique pattern of the antibody binding “distinct...TS (2015) The Economics of Reproducibility in Preclinical Research. PLoS Biol 13(6): e1002165. https://...

  12. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    Published
    Jan. 5, 2021, 2:15 p.m.
    ... Jesse Zalatan’s lab deposited plasmids for a chemically inducible CRISPRa system in yeast is mediated...Find the yeast CRISPRa plasmids here Reduce the scale of your CRISPR screens by using this lentiviral ...paired guides per construct and allows a reduced scale in screens without any reduction in performance ... channelrhodopsin (C1V1) targeting parvalbumin cortical interneurons is now available as a ready to use...

  13. Story of a SynBio Startup: RevBio's Epiphany (or Lack Thereof)

    Type
    Blog Post
    Published
    June 5, 2014, 3:32 p.m.
    ...things more interesting, we are working in a new technical discipline that nobody understands (synthetic ...thing is opening a pizza joint. Silly ideas (mining local salt deposits to sell at farmers markets), and serious...that survived the literature review, survived technical feasibility analysis, and was a marketable product...and rapidly indicate their presence. They are genetically modified organisms, and GMOs have a bad reputation...

  14. Hot Plasmids: Summer 2025

    Type
    Blog Post
    Published
    Aug. 28, 2025, 1:15 p.m.
    ...the ωRNA (TnpB) or a gRNA (Cas9). Drawings not to scale. Created with BioRender.com. Steven Jacobsen...Rodriguez-Fraticelli recently developed an approach called Simultaneous Tracking of Recombinase Activation...recombinase payload activates a floxed reporter gene (genetically encoded or delivered separately). Figure...Cre-dependently to allow tracing exclusively from genetically determined cells. This version of ATLAS works...

  15. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    Published
    June 25, 2024, 1:15 p.m.
    ... Liu, 2023). This method uses a prime editor (typically a Cas9 H840A nickase fused to a reverse transcriptase... design of the pegRNA has been shown to play a critical role in the editing efficiency of prime editing...experimentalist, and especially shines once large-scale prime editing experiments come into play. Nicolas...s41587-023-01678-y Li, X., Chen, W., Martin, B. K., Calderon, D., Lee, C., Choi, J., Chardon, F. M., McDiarmid...

  16. Academic vs. Industry Postdocs

    Type
    Blog Post
    Published
    Nov. 13, 2018, 1:22 p.m.
    ...can be long—5 years or more, especially in the biological sciences—whereas industry postdocs have a set...course, you might have less time to explore scientifically since you can’t afford to spend too much time...Unless your academic lab is closely involved with clinical trials or has ties to industry, the drug discovery...industry postdoc, you might find yourself feeling scientifically isolated if no other postdocs are in your department...

  17. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    Published
    Dec. 1, 2016, 3:30 p.m.
    ...your gene of interest is difficult to target specifically with a gRNA). Be sure to keep these techniques...sequences are exchanged between two similar or identical molecules of DNA. Gene targeting takes advantage...homology is required, but 6 to 14 kb of homology is typical for targeting constructs. In the example shown ...cells participating in development or other physiopathological events to which the chosen promoter responds...

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

    Type
    Blog Post
    Published
    Oct. 23, 2018, 11:49 a.m.
    ...margins of my protocol. While these tips weren’t critical to the experiment, they definitely made my life...qPCR Protocol for more details. Pro Tips When calculating the titer for your AAV, remember to account for...control (NTC) should always be included. While the physical titer generated by qPCR is useful, I test a few...tool for gene expression in mammals Use the PEI calculator for planning AAV packaging transfections Find...

  19. The Scientific Conference Poster Session: Tips for Success

    Type
    Blog Post
    Published
    Aug. 29, 2019, 12:50 p.m.
    ...therapeutic strategies to target it, than you typically would at a microbiology meeting. Create an outline...lab mates, and PI can provide feedback on your technical presentation, but those outside of your field ... through the list at the session so you can strategically use your time. Don’t have access to a list of...titles to find the ones you’d like to revisit. Strategically traverse the sea of posters If you come prepared...

  20. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    Published
    May 29, 2014, 1:29 p.m.
    ... 2017. At Addgene, we continually use the Basic Local Alignment Search Tool (BLAST) provided by NCBI. ...BLAST sequence databases from the dropdown menu. Typically, we use the default nucleotide database “Nucleotide...” program for blastn so that you can extract practically every single reliable base pair from your results... the “Align two or more sequences” option to drastically reduce waiting time for search results. Optimizing...

  21. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    Published
    March 17, 2016, 2:30 p.m.
    ...-length viral genome (Figure 2). While it is technically possible to encode all four viral genes in the... (see Uses of Minigenome System). From a more technical standpoint, a plasmid with all four genes and ... gene by altering the amount transfected (see Technical Tips). Minigenome replication and transcription...compounds which often target the viral polymerase. Technical tips It is important that all of these plasmids...

  22. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    Published
    Oct. 22, 2015, 2:30 p.m.
    ...lab tool. In the classic two hybrid system (also called the interaction trap), genes encoding proteins ...experiments. False positives can occur even in small scale experiments, although these can often be accounted...that the configuration of your fusion proteins physically blocks the binding sites for the protein partner...Tachihara, K., Umezawa, Y.l. Split luciferase as an optical probe for detecting protein-protein interactions...

  23. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    Published
    Oct. 6, 2015, 2:30 p.m.
    ...that, they also help us keep track of the tubes’ physical location in the lab. These barcodes additionally...  freezers depending on the sample type. It is critical for us to know which plate, rack, and freezer ...performs an average of 140 transformations. The lab typically transforms newly deposited samples from Monday...Excel sheet.3. Be specific  - When assigning a physical location to your samples, include as much detail...

  24. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    Published
    Dec. 2, 2014, 7:23 p.m.
    ...been working on development of technologies for biochemical analysis of genome functions such as transcription...researchers to use enChIP in a wide variety of biological systems. We are currently developing several ...Therefore, my lab is dedicated to development of new biological technologies. The locus-specific ChIP technologies... effort to interact with each other, which is critically important, especially for young researchers. ...

  25. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    Published
    July 30, 2024, 1:15 p.m.
    ... ligands (Figure 2). Biosensors, which include calcium and voltage sensors, are the broadest category ...catalog If you’ve been following our blog, you may recall our first 20th anniversary story on CRISPR, where... vectors available in the repository. GCaMP, a calcium indicator (biosensor, by Addgene’s categorization...Waxmonsky, N. C., Swanson, L. E., & Fan, M. (2016). Practical Considerations for Using Pooled Lentiviral CRISPR...

  26. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    Published
    May 2, 2019, 12:41 p.m.
    ...studying parenting strategies and make important ecological indicators since species live in water and on... as the only Monarch butterfly food. They have chemical pathways generating interesting drug-like compounds...Lab (@EvoEmbryo) September 24, 2018   Xenopus tropicalis (Gary Gorbsky) This is not a new experimental...plants and are unusual enough to be interesting biologically. They have economic importance as harmful weeds...

  27. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    Published
    Jan. 3, 2019, 1:35 p.m.
    ... defective gene, Xm, the sequence of which is identical to the correct gene, Xw, except for a single base...precisely changes a single base with an efficiency typically ranging from 25-75%, while the success of precise... in the DNA by cutting only a single strand, dramatically reducing the likelihood of detrimental indel...known 'DNA adenosine deaminases' which would theoretically give rise to an adenine base editor. A massive...

  28. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    Published
    Nov. 1, 2018, 12:35 p.m.
    ...passed down through generations (Figure 1). These so called “plasmid addiction systems” require dividing cells..., plasmid addiction systems have been used as biological tools for a wide range of applications in molecular...become a powerful tool to address a diverse set of biological inquiries. References Ogura, Teru, and Sota...vaccine strain CVD 103-HgR." FEMS Immunology & Medical Microbiology 57.1 (2009): 69-79. PubMed PMID: 19678844...

  29. Transferable Skills Guide: External Collaborations

    Type
    Blog Post
    Published
    April 19, 2018, 1:25 p.m.
    ...gives Snapgene great exposure to the community of biological scientists who use Addgene everyday. To get this...from other institutions personally on conference calls and make sure to maintain these relationships by...speakers for a meetup or recruit volunteers for your local animal shelter. There are many opportunities so ...Universities themselves can help you do this through practical tools like alumni networks. I find it useful to...

  30. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    Published
    Nov. 10, 2016, 3:30 p.m.
    ...possess cell walls. As a result, Mycoplasma are intrinsically resistant to this class of antibiotic. Acquired...When antibiotic resistance testing is requested clinically, this involves growing a standardized amount ...certain antibiotic were chosen for treatment. This is called a Kirby-Bauer disk diffusion test or Kirby-Bauer...-trusted method that is frequently used in the clinical laboratory. What can we do to stop the emergence...
Showing: 511 - 540 of 846 results