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We narrowed to 587 results for: des.1

Showing: 121 - 140 of 587 results

  1. Some Like it Hot: Thermostable GeoCas9

    Type
    Blog Post
    Published
    Sept. 14, 2017, 12:40 p.m.
    ...Figure 1 for a summary of the experimental results. GeoCas9 RNP editing in mammalian cells Besides being...Cells? Yes Yes Longevity in Serum?* No Yes Table 1. Comparing SpCas9 and GeoCas9. *For in vitro Cas9 ...GeoCas9 is active at high temperatures Figure 1: Thermostability of SpCas9 vs GeoCas9. SpCas9 or GeoCas9...from 25-80 °C. Activity is expressed as Kcleave (min-1). SpCas9 has maximum activity from 35-45 °C, while...

  2. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    Published
    Sept. 17, 2019, 12:40 p.m.
    ...production of the pro-inflammatory cytokine interleukin IL-1 beta in the dorsal raphe nucleus (DRN), a region thought... a transgenic mouse harbouring a Cre-dependent IL-1 beta overexpression cassette. Expression of Cre in...in these neurons turns on overexpression of IL-1 beta. They tested the mice in a series of behavioral ...risky), or a closed arm (safe). Mice with higher IL-1 beta production spent more time in the open arm and... part of the apparatus, it turned out that the IL-1 beta overexpressing mice spent more time in the risky...Over the past decades, neuroscientists have made great strides to map brain regions and allocate specific...for this task. However, most lesioning methods destroy areas larger than intended and may unintentionally...

  3. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    Published
    June 22, 2023, 1:15 p.m.
    ...including yourself as the researcher.   Figure 1: Separation of retroviral packaging components onto...these reasons, AAV is usually classified as a BSL-1 safety level. Adenovirus is similar to AAV in that...lab coats are either recommended or required in BSL-1 and required in BSL-2 labs. When working with lentivirus...required whenever there’s a splash or spray risk in BSL-1. In BSL-2 labs, eye protection is required any time...isopropyl alcohol, 0.5% peracetic acid, 10% bleach, or 1% iodine are useful. Some organizations or cities may...that are built into how something functions, were designed for this purpose. One such control is dividing...

  4. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    Published
    April 5, 2018, 12:42 p.m.
    ... Figure 1: In conventional gene targeting (A), you must first generate your desired mutation in mouse...much more favorable than previous methods yielding 1-10% knock-in. Read on to learn how you can make CRISPR... drug induction systems (ttA or rtTA) are between 1 to 2 kb in length. There are plenty of Addgene plasmid...CRISPR ribonucleoproteins.” Genome Biol. 18 (2017) (1):92. DOI: 10.1186/s13059-017-1220-4 PubMed PMID: 28511701... using long ssDNA donors.” Nat Protoc. 13 (2018) (1):195-215. DOI: 10.1038/nprot.2017.153 PubMed PMID:...engineering using CRISPR nucleases. Genome Biol. 19 (2018) (1):25. DOI: 10.1186/s13059-018-1400-x PubMed PMID: 29482575...using in mouse transgenics. Designing an Easi-CRISPR knock-in allele To design a knock-in allele at a given...

  5. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    Published
    Aug. 30, 2022, 1:15 p.m.
    ...Fig. 1). So, while organic solvents may improve antibody binding for some targets, they can destroy it ...preventing your primary antibody from binding (Fig.1). The level of cross-linking that occurs depends on...thing depending on your experiment.   Figure 1: Choice of fixation method impacts antibody binding...New York, pp 299–311. https://doi.org/10.1007/978-1-4939-8935-5_26  Rolls G (2022) Process of Tissue Fixation...www.leicabiosystems.com/us/knowledge-pathway/fixation-and-fixatives-1-the-process-of-fixation-and-the-nature-of-fixatives...critical to the outcome of your IF experiment and deserve careful thought and attention.   * There are other...

  6. 500,000 Plasmids Shipped and Counting!

    Type
    Blog Post
    Published
    March 26, 2015, 3:24 p.m.
    ...here at Addgene – 500,000 plasmids shipped! That's a 1/2 million plasmids sent to happy scientists around...start out at our Cambridge office – previously at 1 Kendall Square, but newly relocated to 75 Sidney Street...Have a question? Email: [email protected] Call: +1 (617) 225-9000         ...information in our lab software, labeling stabs with barcodes as they go. Stabs are organized by shipment and...

  7. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    Published
    Feb. 27, 2018, 2:04 p.m.
    ...incubation of 1° and 2° Must use different species and/or different IgG subclasses of 1° for multi-color...displacement 2° incubation can be skipped by pre-binding to 1° Monovalent Monoclonal Only anti- rabbit and -...dyes are also commercially available. References 1. Pleiner, T., Bates, M., & Görlich, D. (2017). A toolbox...NIH’s Developmental Studies Hybridoma Bank (DSHB). 1 ml of antibody-containing hybridoma supernatant is...their production requires animal sacrifice and precludes genetic engineering of a tag or fusion with a ... with secondary nanobodies conjugated with the desired fluorophores. In figure 4B below, triple colocalization...review. If you just want to use the nanobodies described in Pleiner et al, the key steps for generating...

  8. Clear and Accessible Writing

    Type
    Blog Post
    Published
    Feb. 15, 2024, 2:15 p.m.
    ...writing Let’s use Figure 1 as our example. Remember, our goal is to use words to describe the image in a way...information as sighted readers.    Figure 1: An image from AAV Purification by Iodixanol Gradient...are they hoping to get from it? The image in Figure 1 appears on our protocol page for AAV Purification ... share with the rest of the workshop.  For Figure 1, Addgene chose this alt text: "The left panel is a... to our website through the use of alt text to describe images in online spaces. Alt text is used by screen... your alt text, use all of this information to describe the image helpfully. You and your readers are ...learned a lot from listening to others' alt text descriptions, and we’d love to hear yours as well.  More ...

  9. Quick Guide to Working with Drosophila Part 2: Controlling Gene Expression in Flies with Gal4/UAS

    Type
    Blog Post
    Published
    July 21, 2017, 12:48 p.m.
    ...transcription activation system co-opted from yeast (1). It is a Drosophila geneticist’s main workhorse to...he still reads “#” as a “pound symbol”. References 1. A. H. Brand, N. Perrimon, Targeted gene expression... A Fly Geneticist’s Swiss Army Knife, Genesis 34, 1–15 (2002).PubMed PMID: 12324939. 3. S. E. McGuire,...Versatile Toolkit for Gene Expression in Drosophila, 3, 1–10 (2015). PubMed PMID: 21356876. 5. J. Chow, Z. Marka...references and web links from my first post and start designing your first fly experiment! Many thanks to our...PMID: 28704652. 6. V. Rodríguez, D. Didiano, C. Desplan, Power tools for gene expression and clonal analysis...

  10. Plasmids 101: Mammalian Vectors

    Type
    Blog Post
    Published
    March 25, 2014, 3:15 p.m.
    ...premature chain termination 1-10 ug/mL Zeocin Sh bla HeLa, NIH3T3, CHO, COS-1, 293HEK, Jurkat T cells Complexes...expressing the Epstein–Barr virus (EBV) nuclear antigen 1 (EBNA1) or the SV40 large-T antigen (293E or 293T ...Concentation*** Blasticidin bsd  HeLa, NIH3T3, CHO, COS-1, 293HEK Inhibits termination step of translation ...mammalian plasmid components, it may be useful to describe the means of introducing genetic material (such...the plasmid in the same way. Selection methods, described below, are typically employed when creating stable...

  11. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    Published
    Jan. 26, 2016, 3:30 p.m.
    ... Figure 1: Exon skipping strategies for Duchenne muscular dystrophy. gRNAs can be designed to flank ...X-linked recessive disorder affecting approximately 1 in 5000 males. DMD is caused primarily by frameshift...needed. First, CRISPR delivery must be optimized to: 1. Reach a high percentage of muscle cells throughout...also be closer than we might expect.   References 1. Kaiser J. CRISPR helps heal mice with muscular dystrophy...diseases. Nucleic Acid Ther. 2014 Feb;24(1):69-86. PubMed PMID: 24506781. PubMed Central PMCID: PMC3922311...);186-91. doi: 10.1038/nature14299. Epub 2015 Apr 1. PubMed PMID: 25830891. PubMed Central PMCID: PMC4393360...dystrophin transcript. Unfortunately, the oligonucleotides only modestly improve muscle function, and they must...

  12. Early Career Researcher Toolbox: Social Media for Scientists

    Type
    Blog Post
    Published
    Dec. 11, 2018, 12:46 p.m.
    ...how scientists connect on Twitter that I realized 1) a lot of scientists are using social media professionally... but my consistent writing for the Addgene blog (~1-2 blog posts per month) and use of Twitter and LinkedIn... have tips for using social media as a scientist. 1. "5 Ways Scientists SHOULD Be Using Twitter" podcast...of interest. Hopefully through these links, I’ve: 1) convinced you social media can positively impact ...’s social media toolbox. Besides advice for using Twitter, this post provides suggestions for using LinkedIn...use Twitter as a scientist. While this post is designed with beginners in mind, there’s some great reminders...sheet from Science Media Savvy This tip sheet provides advice for tweeting and blogging as a scientist...

  13. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    Published
    March 25, 2025, 1:15 p.m.
    ...for more details on how they work!) Figure 1: Base transitions are edits between the purine bases...R-loop structure. In order to bind DNA, Cas9 requires (1) a guide RNA, (2) a complementary DNA sequence, and...given the pithier name CGBE1, for “C → G Base Editor 1,” and had a C → G editing efficiency of up to 68%... the CGBE: DNA polymerase D2 (POLD2), exonuclease 1 (EXO1), and RNA binding motif protein X-linked (RBMX...excision repair proteins. Nature Communications, 12(1), 1384. https://doi.org/10.1038/s41467-021-21559-9...transversions in human cells. Nature Biotechnology, 39(1), 41–46. https://doi.org/10.1038/s41587-020-0609-x...machine learning methods. Nature Communications, 12(1), 4902. https://doi.org/10.1038/s41467-021-25217-y...

  14. Antibodies 101: Choosing the Right Isotype

    Type
    Blog Post
    Published
    April 6, 2023, 3 p.m.
    ..., rather than a broadly  reactive secondary (Fig. 1). For example, an anti-mouse IgG2a secondary would...IgG2a>IgG2b>IgG1 (Manning 2012).      Figure 1: Broadly reactive versus subclass-specific antibodies...antibody binding and therefore the signal strength (Fig. 1). Monoclonal primaries If you do decide on a monoclonal...raised in a different species.      Table 1: Isotype considerations when choosing antibodies ...Elsevier. https://doi.org/10.1016/s0091-679x(08)60255-1  Manning, C. F., Bundros, A. M., & Trimmer, J. S. ... and to work in a variety of applications. The desired specificity of your secondary antibody depends ...

  15. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ...journal publication describing the plasmids, or (for deposits submitted after July 1, 2025) after three... emailed submission) in the Deposit menu: Figure 1: Begin the deposit process by clicking "Submit Online...cases Please contact us at [email protected] or +1 (617)-ADDGENE (233-4363) if your deposit falls into...Addgene?" or contact us at [email protected] or +1 (617)-ADDGENE (233-4363) with additional questions...strain. Please contact us at [email protected] or +1 (617)-ADDGENE (233-4363) if you’d like to distribute...page so that any data you enter is not lost.   Step 1: Sequences, Maps, and Files Please provide any full...a 1.5 mL microfuge tube at a concentration of 0.1–1 µg/µL (or bacterial streaks, if you prefer). If you...

  16. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    Published
    Sept. 12, 2017, 1:44 p.m.
    ...Updated Jun 1, 2021 by Meghan Rego. Reproducible data are key to science, so scientists are used to repeating...the DNA packaged inside the viral particles (Figure 1). Briefly, packaged DNA is extracted from purified.... When contamination is suspected, we proceed to de novo assembly of the unmapped reads, employing our...transduced cells whenever possible. This post will describe our workflow for viral genome sequencing (VGS)...

  17. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    Published
    June 18, 2020, 1:15 p.m.
    ...crevices proteins on the cell surface. Figure 1: Comparison of common antibody, HCab, and nanobody...Humana Press, pp 133–148. https://doi.org/10.1007/978-1-62703-293-3_10 Walter J et al. (2020) Synthetic nanobodies...https://doi.org/10.1101/2020.04.16.045419.  Wrapp D, De Vlieger D, Corbett KS, Torres GM, Wang N, Van Breedam...Zimmermann et al., 2020). This selection platform was designed so that any standard lab can quickly select for...strongly to your target protein. The Seeger lab designed 3 different mRNA sybody libraries for use in the... randomized with a defined set of different trinucleotides (corresponding to different amino acids) in...for binding to the target protein. The protocol describes going through two rounds of phage display to increase...

  18. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

    Type
    Blog Post
    Published
    Sept. 5, 2017, 1:58 p.m.
    ...recently that desired genome-editing events by HDR can be generated more efficiently by (1) improving editing...specifically incorporate homo- or heterozygous mutations. 1. Increasing HDR accuracy with CRISPR/Cas-blocking ...targeted alleles when gene editing.   Figure 1: Techniques to improve editing efficiency.   But...provided by blocking mutations. Essentially, you:  Step 1 - Introduce the blocking mutation together with the...developing novel iPSC-based model systems.     References 1. Cong, Le, et al. "Multiplex genome engineering using...genome editing with CRISPR/Cas9 in human cells was described in 2013 (Cong et al, Mai et al) we were very excited...introduced. Although we were able to introduce the desired mutations in genes associated with dementia, such...

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

    Type
    Blog Post
    Published
    Oct. 6, 2015, 2:30 p.m.
    ...necessary.   Tips for barcoding and tracking samples 1. Start with the tube - Barcoded tubes come in many...all of these samples have the 2D barcodes mentioned above. These barcodes identify what type of sample is...tubes. 1D barcodes are better for tubes that are meant to be read one at a time. 1D barcodes can be purchased...the tubes’ physical location in the lab. These barcodes additionally allow us to store multiple types ...fresh media with antibiotics at each stage. The barcodes and LIMS help us track each sample, but we also... a petri dish plate that is also barcoded. The barcodes on both of these are scanned into our LIMS system...samples within your lab, we highly recommend using barcodes! Barcoded tubes are easily accessible and can ...

  20. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    Published
    May 14, 2024, 1:15 p.m.
    ...Combinatorial Design for Synthetic and Traditional Biology.” ACS Synthetic Biology, vol. 5, no. 1, American...within the mitochondrial matrix.     Figure 1: A chart with MoClo kit recommendations for different..., vol. 4, no. 9, American Chemical Society (ACS), 1 May 2015, pp. 975–86. Crossref, https://doi.org/10.1021...Yeast Mitochondria.” BMC Biotechnology, vol. 17, no. 1, Springer Science and Business Media LLC, 13 Nov. ...Editing in Plants.” BMC Plant Biology, vol. 20, no. 1, Springer Science and Business Media LLC, 23 Apr. ...Arabidopsis Mutants.” The Plant Journal, vol. 106, no. 1, Wiley, 25 Mar. 2021, pp. 8–22. Crossref, https://...Cloning Platform.” Scientific Reports, vol. 9, no. 1, Springer Science and Business Media LLC, 12 July ...
Showing: 121 - 140 of 587 results