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We narrowed to 672 results for: abo.1

Showing: 221 - 240 of 672 results

  1. Multiplexed Overexpression of Regulatory Factors (MORF) collection

    Type
    Blog Post
    ...the viral TF library, with each cell receiving only 1 TF expression construct. Cells were cultured following...states was generated (Joung et al., 2023). Fig. 1: Barcoded TF library was packaged in lentiviral particles...atlas of directed differentiation. Cell. 2023. 186 (1) 209-229. doi: 10.1016/j.cell.2022.11.026 Joung J....changes. Using this road map, and what was known about previously characterized TFs, the Zhang group were...Additional Resources on the Addgene Blog Learn More about Lentivirus Figure out How to Get the Most from Your...

  2. Flow Cytometry Readouts: Yes, No, and Everything in Between

    Type
    Blog Post
    ...overlap with the negative control population (Figure 1). Based on where the GFP- population ends, you can...total cells present in the gate.   Figure 1: Example of yes-no flow plot of the (negative) control...for a yes-no dot plot, like the one shown in Figure 1.  Figure 3: An example of a histogram plot ...interest. Created with biorender.com. The plot above is for a smeary expression and is quite different.... Additionally, you can gate off the histograms above, allowing you to visualize your data as a dot plot...

  3. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    ... is conjugated to a fluorophore (Figure 1).   Figure 1: Comparison between how direct immunofluorescence...important considerations are to make sure your secondary 1) will recognize the species of your primary antibody...New York, pp 299–311. https://doi.org/10.1007/978-1-4939-8935-5_26 Stuurman N. (2012) Introduction to ... Learn about research applications of antibodies in Introduction to Antibodies Learn more about fluorescence...Stay tuned for a future blog post to learn more about these techniques!  Direct vs. indirect immunofluorescence...different approaches to be familiar with when talking about IF, direct and indirect. For direct IF, researchers...immunofluorescence procedure with key considerations about each step. Image created with BioRender.com ...

  4. Hot Plasmids: Winter 2025

    Type
    Blog Post
    ...new combinations of PE protein components (Figure 1), an altered epegRNA delivered with an enhanced pU6cm...amplification of cargo, and heat treatment.  Figure 1: Left: recombined prime editing tools PE2max-NC and...conformational dynamics of the motor protein kinesin-1, the Twelvetrees Lab took advantage of the self-labeling..., Craggs, T.D., & Twelvetrees A.E. (2024) Kinesin-1 is highly flexible and adopts an open conformation...synaptic events By Brian O’Neill The Lin Lab and collaborators recently created ASAP5, a faster and more responsive...

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

    Type
    Blog Post
    ...required for replication and transcription (Figure 1). All viral genes are removed and replaced by a single...cells that stably express T7 (e.g. BSR T7/5 cells) (1). We hope that minigenomes will greatly enhance our...transcription of negative strand RNA viruses.   References 1. Buchholz, Ursula J., Stefan Finke, and Karl-Klaus...reverse genetics. Springer Berlin Heidelberg, 2004. 1-41. PubMed PMID:15298166. 3. Hoenen, Thomas, et al...Microbiology, National Emerging Infectious Disease Laboratories.  She is interested in genome replication and...

  6. Chemogenetics Guide

    Type
    Guide
    ...out our Chemogenetics Plasmid Collection . Figure 1: General overview of a chemogenetics experiment. Early...bidirectional control of neuronal activity. Table 1. Descriptions of DREADDs and their activity in neurons... while PSAMs and LMOs have an effect for only 0.5–1 hours after delivery. Chemogenetic ligand. The chemogenetic.... However, CNO is back-metabolized to clozapine and other clozapine metabolites and these can have off-target...concentrations of clozapine N ‐oxide (CNO). CNO is a metabolite of the antipsychotic clozapine and seems to be...experiments in this species may be entirely due to back-metabolism of CNO to clozapine, demonstrating the need for...activity, and do not seem to have the same back-metabolism issues as CNO and are attractive alternatives...

  7. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ... and reproducibility of a study’s findings. Table 1: Biased Genotyping Options Technique Sequence Information...sequencing doesn’t tell the whole story Figure 1: Schematic representations of the indicated unbiased...types Must be verified with cell-based method IDLV 1% Cell-based Programmable, can detect DSBs in live ...magazine.scienceforthepeople.org/.     References 1. Fu, Yanfang, et al. "High-frequency off-target mutagenesis...next-generation sequencing." BMC genomics 15.1 (2014): 1. PubMed PMID: 25409780. PubMed Central PMCID: PMC4246457...et al. 2015). Meanwhile, Sanger sequencing is laborious, time consuming and cannot be applied to heterogenous...viable options for multiplex sequencing in many laboratories. Nevertheless, unbiased detection isn’t as sensitive...

  8. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...10.1038/s41551-024-01227-1. doi:  https://doi.org/10.1038/s41551-024-01227-1   CHARM: A compact epigenetic...spacer proteins ending with a GFP nanobody (Figure 1). This enrichment streamlines the purification and...highly-visible beads on the cryo-EM grid.     Figure 1: MagIC-cryo-EM for sample enrichment and structure...single-particle analysis of PRC2 with RvLEAMshort (1:6 molar ratio) with 10 minutes of glutaraldehyde crosslinking...transcriptional silencing (Nuñez et al., 2021). Now, in a collaboration with the Vallabh/Minikel Lab, the Weissman Lab...

  9. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...example, Cas9 from Staphylococcus aureus (SaCas9) is about 1 kilobase smaller than SpCas9, so it can be packaged... modify any desired genomic target provided that (1) the sequence is unique compared to the rest of the...will cover three ways to circumvent this limitation: 1) the use of novel S. pyogenes Cas9 variants with varying...Biol 19: . https://doi.org/10.1186/s13059-018-1591-1 Cox DBT, Gootenberg JS, Abudayyeh OO, Franklin B, ...Resources on the Addgene Blog  Read more about Cpf1 Read more about SaCas9 and AAV Check out our CRISPR Featured... and VRER) recognize novel PAM sequences (shown above). The VQR, EQR, and VRER Cas9 variants are capable...

  10. Form Your Own Peer Mentoring Group: A How-To Guide for Scientists

    Type
    Blog Post
    ... experimented with a group mentoring format where 1 mentor met with a group of mentees to get more “bang...The groups in these programs typically consist of 1-2 mentors and 3-5 mentees who meet together for an...get together. If you all work at the same place, a 1-2 hour lunch on a regular schedule might work. If ...for Scientists Series. I have been thinking a lot about Mentoring for over 10 years. Many successful scientists...Medical School researcher Connie Cepko to learn about her mentoring style. While working on this project...postdocs I have started to talk to trainees more about peer mentoring. They want to find mentors to help... questions I get is, “What are we going to talk about?” There are an infinite number of ideas on the internet...

  11. Reagent Repositories Are Speeding up Science During the Pandemic

    Type
    Blog Post
    ...transmissibility, and laboratory studies of COVID-19: an observational study. The Lancet Microbe 1:e14–e23 . https...available from their original sources. Figure 1: Biological repositories that distribute mouse strains... from Reproducibility for Everyone. Case study 1: Cell lines Many of the reagents used to investigate...the Addgene blog Read more about COVID-19 databases and resources Learn about Addgene's work to share plasmids... availability of this strain from The Jackson Laboratory. Originally developed and characterized by Stanley...mouse strain is regaining popularity. The Jackson Laboratory, a nonprofit research institution, has been able...

  12. Viral Vectors 101: Viruses as Biological Tools

    Type
    Blog Post
    ...unleashes its cargo – its own genome.   Fig. 1: Viral particle infection into a host cell. Image ...Vector Resources at Addgene! References and Resources 1. Roe T, et al. “Integration of murine leukemia virus...nuclear import of human immunodeficiency virus type 1 preintegration complexes.” Proc. Natl. Acad. Sci. ...applications to help you start (safely!) thinking about the possibilities. Ready to plan your next (or maybe...S, Varmus H. Retroviruses. Cold Spring Harbor Laboratory Press; Cold Spring Harbor, NY, USA: 1997. PubMed...Additional Resources on the Addgene Blog Learn More about Lentivirus Listen to Connie Cepko Discuss Gene Therapy...Addgene.org Browse Our Viral Vector Pages Learn About Viral Vector Biosafety Deposit Your Viral Vectors...

  13. Visualizing Your Plasmid Request Data

    Type
    Blog Post
    ...Figure 1: The Addgene Requests Dashboard   The dashboard will display the following items (Fig. 1): ...analyzing and sharing your own depositor metrics!  About the Author Joachim Goedhart is an assistant professor...

  14. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...indicates the presence of a multimer.   Fig. 1: The monomer of Plasmid 12259: pMD2.G (depicted on...plasmid sequencing services using long-read sequencing (1-25 kb), such as Oxford Nanopore sequencing or PacBio...10017-26. doi: 10.1074/jbc.M110.176636. Epub 2011 Feb 1. PMID: 21285356; PMCID: PMC3060453. Bazaral M, Helinski...multiple circular DNA forms of colicinogenic factor E-1 from Proteus mirabilis. Biochemistry. 1968 Oct;7(10...Methods Mol Biol. 2009;582:11-25. doi: 10.1007/978-1-60761-340-4_2. PMID: 19763938. Maucksch C, Bohla A...colonies using one of the detection methods described above can help ensure the isolation of the monomeric version...plasmid DNA production platform incorporating low metabolic burden seed-stock and fed-batch fermentation processes...

  15. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ...between 2 loxp sites and is excised by Cre (Figure 1). This results in one loxp site and one lox2272 site...cause of this spurious expression?   Figure 1.  Cre-recombinase binds to either pair of lox sites...recombination and can exhibit recombination levels as high as 1-3%. In light of this finding, we now include guidance...Efficiency, Tropism and Axonal Transport of AAV Serotypes 1, 2, 5, 6, 8 and 9 in the Mouse Brain. PLoS ONE 8:e76310...Vectors Pseudotyped with Viral Capsids from Serotypes 1, 2, and 5 Display Differential Efficiency and Cell...the most common concerns we get here at Addgene about this system is off-target or spurious expression...cells that don’t express Cre or Flp, what can we do about it?  Test the serotype and promoter The first step...

  16. Antibody Neutralization Response Against Pseudoviruses Expressing SARS-CoV-2 Spike Protein Variants

    Type
    Blog Post
    ...for the wild-type. However, variants such as the P.1 (Gamma variant) and the B.1.351 (Beta variant) were... tested at the time of this study. Figure 1: Neutralization levels against spike pseudovirus decreases...lowest levels of cross-neutralization occurs in the P.1 and B.1.351 variants tested. Image from Garcia-Beltran...Browse all COVID-19 related blog posts Learn more about pseudotyping Resources on Addgene.org Find plasmids...

  17. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

    Type
    Blog Post
    ... you’d like to use to generate your new fly line: 1. Where would you like the gene to be incorporated ...control exactly in which locus your transgene ends up (1): Random insertion has the advantage that you can...he still reads “#” as a “pound symbol”. References 1. J. R. Bateman, et al. Site-Specific Transformation...immunofluorescent images like in this paper (2). Metabolic assays such as Seahorse can be employed to look...Additional Resources on the Addgene Blog Learn about Using Cre in Drosophila Read Other Genome Engineering...

  18. Screening for Successful Genome Editing with Digital PCR

    Type
    Blog Post
    ...digital) signal at the end of the reaction (Figure 1). Partitions that did not contain any target DNA at...integrated through homology-directed repair (HDR) (1), or indel mutations resulting from non-homologous...Follow him on Twitter @AnotherLabRat.     References 1. Miyaoka, Y. et al. Isolation of single-base genome-edited...advantages over mismatch nuclease assays that will be elaborated below. Digital PCR Digital PCR (dPCR) involves...type targets. Comparatively, detection limits of about 5% mutant targets have been reported for mismatch...

  19. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    ...visualizing hM4D(Gi)-mCherry in fixed tissue. References 1. Aschauer DF, Kreuz S, Rumpel S. Analysis of transduction...efficiency, tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PLoS One. 2013...PMC3069789. 3. Kügler S, Kilic E, Bähr M. Human synapsin 1 gene promoter confers highly neuron-specific long-...Resources on the Addgene Blog Learn More About Lentivirus Learn more About AAV Get the Scoop on Our Viral Service...

  20. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    ...Collection Fluorescent tagging strategy Figure 1: Gene editing is the addition of a useful coding sequence...plasmid design strategy. A donor plasmid contains 1 kb homology arms matching the gene of interest, a ...Institute website at alleninstitute.org. References 1. Roberts, Brock, et al. "Systematic gene tagging using...the Addgene Blog Learn More about Homology Directed Repair Learn More about Non-Homologous End Joining .... For a C-terminal tag, as shown in the example above, we inserted a linker and GFP tag at the end of ... and 1kb of homologous DNA flanking both sides (about 2.7kb total)—is inserted into a plasmid backbone... Plasmids, plasmids everywhere To find out more about gene edited hiPS cell lines generated using the ...
Showing: 221 - 240 of 672 results