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Showing: 541 - 570 of 954 results

  1. Immunocytochemistry

    Type
    Protocol
    ...PBS, remove the wash, and dispose of it in an appropriate waste container. Fix each well with 500 µL of...as methanol or acetone may be better for some applications. Remove the paraformaldehyde and follow your...safety guidelines for disposing of waste in the appropriate container. Wash 3x for 5 min in 500 µL PBS on...permeabilization buffer and dispose of it in an appropriate waste container. Wash 3x for 5 min in 500 µL ... the blocking buffer and dispose of it in an appropriate waste container. Dilute the primary antibody ...the primary antibody and dispose of it in an appropriate waste container. Wash 3x for 5 min in 500 µL ...min at RT in the dark. Pro-Tip The plate can be wrapped in foil to block light. Remove the secondary antibody...

  2. An Introduction to Adenovirus

    Type
    Blog Post
    ...which a vaccine is available. However, here it appears that the culprit for these unusual cases was either... the removal of certain adenoviral genes that apparently contribute to virulence (Tollefson et al., 1996...on the Addgene blog Viral Vectors 101: Viral Applications Adenoviral Vector Production and Troubleshooting...

  3. Tips for arabidopsis transformation

    Type
    Blog Post
    ...Addgene  Choosing an expression system Choosing an appropriate expression vector is the first step in generating...binary gateway vectors and can be used for promoter swapping and generating N and C terminally tagged fusion...phenotypes. For most phenotypic analysis it is appropriate to begin working in the T2 generation. The selection...

  4. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    ..., Alexa Fluor 568, or Alexa Fluor 647 before application to the sample. Staining with this multi-color...Mouse IgG2a TP1129 pTP1005 (3x cysteines) Mouse Kappa TP1170 pTP1174 (1x cysteine) pTP1172 (3x cysteines...labeling for super-resolution imaging, rapid epitope-mapping and native protein complex isolation. eLife. PubMed...

  5. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    ...What the Future Holds and the Scientists Making It Happen, is an exercise in restraint. The very first subheading... primarily on fluorescent proteins and their applications, tackles much broader issues in this brief volume...ago I would have nodded my head knowingly and appreciatively throughout this chapter. Now, these observations...

  6. A Guide to Starting Your Own Journal Club

    Type
    Blog Post
    ...the most important is letting people know it’s happening. Aim for 3-4 weeks advance notice for the first...the Addgene bioinformatics journal club! What happens during journal club? There are many common ways....  These variations are suited for different applications and should be carefully chosen for each group...

  8. Summer SciComm Series: Modes of Communication

    Type
    Blog Post
    ... or burn yourself out trying to slog through unappealing work in your precious free time.  And don’t be...- will be required to produce an outcome you’re happy with. Know if you’ll need specialized equipment ...then pick whichever is most accessible or most appealing to you. And if you’re not sure what all the possibilities...

  9. AAVs CREATed for Gene Delivery to the CNS and PNS

    Type
    Blog Post
    ... an ideal way to deliver AAV to the PNS. It’s a happy accident that AAV.PHP.S targets the PNS since it...jumble of fluorescent neurons. A simple but flawed approach to multi-color labeling is to inject a mix of ...morphologic studies), as well as translational applications (mechanistic studies and gene therapy treatments...

  10. Plasmids 101: Colony PCR

    Type
    Blog Post
    ...primers. Find Empty Backbones at Addgene Each approach has advantages and drawbacks, which are summarized...which method you choose, make sure to use the appropriate antibiotic for selection.   Lysing bacteria...to determine their size. Make sure to use an appropriate molecular weight standard for reference and to...

  11. CRISPR Antimicrobials

    Type
    Blog Post
    ...virulent or antibiotic resistant bacteria, and inappropriate antibiotic usage selects for resistant bacteria...to combat antibiotic resistance using multiple approaches. Major restaurant chains including Chipotle and...bacterial host range. To further potential clinical applications, Ando et al. recently engineered synthetic phages...

  12. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...or growth stage). A prey library can either be applied to the bait in a single population (e.g. transforming...although these can often be accounted for using appropriate replicates and controls (see Tips below), and.... Obrdlik, P., El-Bakkoury, M., Hamacher, T., Cappellaro, C., Vilarino, C., Fleischer, C., Ellerbrok, ...

  13. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ..., Zetsche et al. developed a Cpf1 multiplexing approach using a single crRNA array to express up to 4 ...-target editing in a cell culture model. The application of Cpf1 to genome editing is exciting both in...in terms of basic science and translational applications. This discovery of this type V CRISPR system ...

  14. The Importance of a Fun Workplace: Company Culture at Addgene

    Type
    Blog Post
    ... result seems to be a balance of productive and happy employees. The teambuilding culture helps us build...throughout the years. This team building merriment happened naturally when Addgene was a family affair back... sure people know that there is something fun happening! Addgene actually has chimes outside of our break...

  15. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    ...which helps increase cellular uptake, but this approach requires compatible cell lines that express viral...individual analysis. A shuttle vector designed for this approach in a mammalian cell experiment should still include...terms are typically used in different (though overlapping) ways. Broad host range plasmids are those that...

  16. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...Paul Blainey’s lab has developed a multiplexing approach to Cas9-based pooled CRISPR screens called CROPseq-multi...for microglia research and broader therapeutic applications.  Find microglia-targeting AAV plasmids here...reconstituting coding sequences for a range of applications, including CRISPR activation by dCas9-VPR, prime...

  17. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    ... the bacteria were injected, limiting their applicability. They also did not scatter ultrasound nonlinearly...chromatic-opening agents to boost expression to appreciable levels. We wanted to create second-generation...ultrasound imaging of nanoscale acoustic biomolecules. Appl. Phys. Lett. 110, 073704 (2017). Hurt, R. C. et ...

  18. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    ...institutions, or institutions they're thinking about applying to, are doing - for example, your benefits may...get rid of postdocs; that current postdoctoral appointments may shorten in length; that postdoc salaries...trainee workforce has clearly not been a sustainable approach to biomedical research in the U.S. and postdoctoral...

  19. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    ...used to generate AAV.  One study showed that approximately 50% of viral capsids were genome-containing ...particles can be identified after negative staining and appear darker than full vector particles. This image shows...experiments? Because (physical) AAV titers are approximations and they depend highly on the experimental ...

  20. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ... its reintroduction, inversion and re-inversion happen at the same probability. Continuous inversion of...the same time from two different plasmids this approach becomes problematic as it reduces the probability... Cell Types for Imaging and Long-Range Circuit Mapping.” Journal of Neuroscience 28(28): 7025–30. PubMed...

  21. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    ...highlights some of the latest breakthrough therapeutic approaches. What is Rett Syndrome? Caused by a mutation ...technical challenges to overcome for both of these approaches, such as delivery of the editing machinery to...Extends the Survival of MeCP2-Null Mice without Apparent Toxicity after Intracisternal Delivery. Molecular...

  22. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ...movement in an open field box or a circular corridor apparatus using a camera or tracking software. Locomotion...and might open new ways for a novel therapeutic approach against drug addiction. Anxiety tests Rodents...measuring the time spent in a particular part of the apparatus, it turned out that the IL-1 beta overexpressing...

  23. The Challenges of Cell Culture

    Type
    Blog Post
    ...especially with industry. Today we also have the appropriate tools to verify our cells lines. Even if you ...the results are reliable. With this two-pronged approach, the many mistakes of the past can be prevented...including GSK, AstraZeneca-MedImmune, and Infors & Applikon. Due to his high level of experience he now travels...

  24. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    ... barrier of the endothelium, but they could be applied to study other actin-dependent processes in cells...) and to introduce unnatural head groups for applications like fluorescent labeling via click chemistry...constructs are great for in vitro biocatalytic applications where toxicity doesn’t matter. Plus, for folks...

  25. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ... explain how to verify that your cells were appropriately edited for your desired mutation - insertion...frequency is 50%, then you could infer that approximately 25% of cells are likely -/-, 50% are -/+, and...sequencing confirmation. All knock-ins A method applicable to knock-ins both large and small is a software...

  26. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    ...study its function is one of the most popular applications of fluorescent proteins. These fusion proteins...targeted to the endoplasmid reticulum based on the appearance of OSER whorls. This assay is quantitative and...linker sequence short In the early days of GFP applications, many were concerned with steric hindrance by...

  27. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...enjoy a warm meal on a winter day, you should be happy to learn that many DNA-based reagents are named ...aptamer soup by exploring the development and applications of fluorescent RNA aptamers in Addgene’s collection...fluorescence when bound to DFHBI despite being approximately half the length (Warner et al., 2014); its length...

  28. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...References Espah Borujeni, Amin, Anirudh S. Channarasappa, and Howard M. Salis. "Translation rate is controlled...Scarless Modification of Proteobacterial Genomes." Applied and environmental microbiology (2019): AEM-00990...chromosomal gene replacement in Escherichia coli." Appl. Environ. Microbiol. 81.15 (2015): 5103-5114. PubMed...

  29. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...polycistronic E. coli expression constructs. Some applications of plasmids from this kit include the production...can be engineered to bind specific RNA bases. By swapping out these different modular repeat segments, you...variety of functions via different fusions. Some applications of PUF protein fusions reported in the literature...

  30. Pouring LB Agar Plates

    Type
    Protocol
    ...concentration disolved in the appropriate liquid solvent. See table to the right for appropriate antibiotic concentrations...Strains Grow Assuming the appropriate strains were streaked on the appropriate plates, then if both strains... LB-agar powder you’ve measured out into an appropriately sized bottle for autoclaving. We make 400 mL...autoclave. Be sure to check the literature for an appropriate sterilization technique if you are working with... wipe down with a paper towel. Count out the appropriate number of plates and stack them on your lab bench...of molten gel mix, a tube rack containing the appropriate antibiotics, and a section for active pouring...antibiotic. Incubate both plates overnight at the appropriate growth temperature and check for growth. See ...
Showing: 541 - 570 of 954 results