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We narrowed to 234 results for: Anti-CRISPR

Showing: 201 - 220 of 234 results

  1. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    ...Biology with BioBuilder 3D Models for learning about CRISPR Resources at Addgene.org Find plasmids from the...riboprobe binding (via hybridization) specific antibodies that detect the mRNA-riboprobe complex are added...stain the cells that contain this mRNA-riboprobe-antibody complex and, as a result, these cells turn blue...

  2. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    ... RNA amplification, new toehold switch designs, CRISPR/Cas9-mediated genotyping, as well as our electronic... synthetic gene networks for the detection of antibiotic resistance and strain-specific diagnostics for...electronic reader that provides portable quantification of diagnostic results.  Our approach now detects samples...

  3. Guide to Using Pooled Libraries

    Type
    Guide
    ...Using CRISPR/Cas9 Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening Pooled CRISPR Libraries... mechanisms for lentiviral CRISPR libraries include: resistance to the anti-cancer drug vemurafenib , ...protocol. Read our CRISPR guide for more information on genome-wide screening with CRISPR/Cas9 . Browse our...virus, make virus; this creates a pooled lentiviral CRISPR library. Apply pooled library to cells. DIVERGE... plasmids have been depleted by selection. In a CRISPR screen, negative screens are often used to identify...

  4. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ... genes for proteins in these compartments using CRISPR systems should strongly consider using moxFPs over...by guest bloggers Erik L. Snapp and Lindsey M. Costantini. "You underestimate the power of the Dark Side...non-fluorescent, and misfolded FP-fusions on quantitative imaging experiments and cell physiology in general...population is not readily apparent and can confound quantitative imaging experiments or even negatively impact...misfolded dark FP fusions including erroneous quantitation of fusion protein levels. It remains unclear...proteins. Our recent publication illustrated the quantitative increase in fluorescent signal achieved when...our guest bloggers Erik L. Snapp and Lindsey M. Costantini! Erik Lee Snapp received his PhD in Molecular...

  5. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    ...- a primer for beginners Adenoviral delivery of CRISPR/Cas9 5 tips for troubleshooting viral transductions...formation assay on A549 cells . Alternatively, antibody-based assays are commercially available for faster...ratio for the purified virus should be ~1.3 Quantitative PCR: the number of viral DNA packaged in virions...virions is determined by a standard curve of known quantity and primers specific for a viral DNA sequence....

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

    Type
    Blog Post
    ...packaging transfections Find tips for a first time CRISPR user Resources on Addgene.org Learn how Addgene...titration by qPCR Overview: AAV titering by qPCR quantifies the number of genome-containing viral particles... primer/probe set can be used and samples are quantified by comparing to an AAV standard curve, which ...Universal real-time PCR for the detection and quantification of adeno-associated virus serotype 2-derived...

  7. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ...page Find synthetic biology plant plasmids Find CRISPR plant plasmids ...track, we switched to luciferases to more easily quantify the effect of the different treatments on transformation...transformation success. Luciferases are great quantitative reporters and use a substrate to produce bioluminescence..., Marc-André, et al. "Transient expression of antibodies in plants using syringe agroinfiltration." Recombinant...

  8. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ... References 1. Ma, Hanhui, et al. “Multicolor CRISPR labeling of chromosomal loci in human cells.” Proc...advantages over their amino acid-based counterparts, antibodies and enzymes, such as: Design and selection ...commercial synthesis (those of us who have used antibodies for diagnostics know how frustrating it can be...hybridization (FISH) where fluorophore-linked antibodies are used to visualize RNA. Low signal-to-noise...

  9. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    ...for transfer plasmids with small inserts (such as CRISPR guides) while a single 72h harvest is best for ...generally a good place to start. If you don’t anticipate that you will need large volumes of lentivirus...

  10. Tips for arabidopsis transformation

    Type
    Blog Post
    ...Addgene blog Engineering the plant genome using CRISPR/Cas9 Using pSiM24 for simplifying plant genetic...the tDNA insertions (usually via seed growth on antibiotic-containing media) A similar process occurs ... the absence of selection. For a protocol for antibiotic selection, check out Harrison et al., 2006. Tips...

  12. Plan Your Experiment

    Type
    Guide
    ... Science Guides CRISPR Guide Plan Your Experiment Plan Your CRISPR Experiment Jump To: Plan Your Experiment...you started using CRISPR in your research. Although we will use the example of CRISPR/Cas9 in mammalian...express the CRISPR machinery. For more difficult cells (e.g. primary cells), viral delivery of CRISPR reagents...conducting genome-wide screens using CRISPR AAV transduction CRISPR elements are inserted into an AAV transfer...lipids Transient expression of CRISPR components Short window of CRISPR activity may decrease off-target...Expression System Design gRNA Publications Get Started CRISPR is a powerful system that enables researchers to... cells, many of these principles apply to using CRISPR in other organisms. First, consider the genetic...

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

    Type
    Blog Post
    ...revolution in genome editing methods (most notably CRISPR/Cas mediated HDR) enables the tagging of endogenous...Organized Smooth Endoplasmic Reticulum (OSER) assay (Costantini et al., 2012) measures homodimerization of FPs...the appearance of OSER whorls. This assay is quantitative and is currently the standard for assessing ...: 30783202. PubMed Central PMCID: PMC6381139. Costantini, Lindsey M., et al. "Assessing the tendency of...PMCID: PMC3324619. Cranfill, Paula J., et al. "Quantitative assessment of fluorescent proteins." Nature ...

  14. Plasmids 101: Biotinylation

    Type
    Blog Post
    ... interactions Learn how biotin can be used with CRISPR Additional resources ThermoFisher Scientific... and antibody labelling. In these experiments, biotin is added to a primary or secondary antibody that...that targets your protein of interest or primary antibody respectively. Avidin bound to a reporter is then... on this complex can bind to your biotinylated antibody. This creates a large meshwork of biotin-avidin...

  15. Evolution of Lab Techniques

    Type
    Blog Post
    ... instance, the popular genome-engineering tool, CRISPR, may one day make techniques used to generate transgenic...from their evolution. Southern blotting and quantitative real-time PCR Southern blotting, named for its...commonly used. In all cases, the result can be quantified by the strength of the signal in comparison to...experimental questions. Real-time PCR detects and quantifies a gene or DNA sequence of interest by recording...Frank Speleman, and Jo Vandesompele. "Real-time quantitative PCR as an alternative to Southern blot or fluorescence...

  16. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...genomic loci in living human cells by an optimized CRISPR/Cas system." Cell 155.7 (2013): 1479-1491. PubMed...N-terminus, but this hardly reduces its size. The substantial size of GFP may interfere with the activity or...: PMC5385933. 6. Cranfill, Paula J., et al. "Quantitative assessment of fluorescent proteins." Nature ...PubMed Central PMCID: PMC3918502.  14. Chernov, Konstantin G., et al. "Near-infrared fluorescent proteins...

  17. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...CRISPR-Cas9 genome editing may be the hot new way to manipulate gene expression, but other gene manipulation...sequences are highly homologous, preventing design of antibodies specific to each fluorophore. Third, Brainbow...mKate2.) They then successfully generated custom antibodies to each of these proteins and confirmed that ...

  18. Addgene's eBook Collection

    Type
    Guide
    ...Close About Overlay Select CRISPR 101 January 2021 | Third Edition About CRISPR 101 January 2021 | ...Overlay Select CRISPR 101 en español January 2024 | First Edition, trans. About CRISPR 101 en español...nuestro libro electrónico, CRISPR 101. Close About Overlay Select Antibodies 101 New! October 2024 | ...experimental advice such as tips for planning your CRISPR experiment and designing your gRNA and how to use...| First Edition About Antibodies 101 October 2024 | First Edition About Our newest eBook contains...contains practical introductions and helpful tips for antibody-based applications, from flow to IHC! Close About...

  19. Science Guides

    Type
    Guide
    ...Read More CRISPR Class 2 C lustered R egularly I nterspaced S hort P alindromic R epeat (CRISPR) systems...background information on CRISPR and the basics for planning your first CRISPR experiment. Read More Optogenetics... below. Antibodies Chemogenetics CRISPR Optogenetics Pooled Libraries Viral Vectors Antibodies Produced...system, antibodies are proteins capable of highly specific binding to other proteins. Antibody-based applications... often use antibodies conjugated to a signaling molecule, can be used to capture, quantify, and identify...

  20. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    ...Knockin Plasmids to Disrupt Genes in Mammalian Cells CRISPR Methods for Bacterial Genome Engineering Resources... system to replace a gene of interest with an antibiotic resistance cassette.   Substrate DNA Design..., including selectable DNA fragments, such as antibiotic resistance genes, as well as non-selectable DNA...lambda red expression is that it doesn’t require antibiotic selection to maintain expression of the recombineering...the cI857 repressor to regulate expression. An antibiotic can be used to select for positive clones but...Selection and confirmation of recombinant clones If a antibiotic resistance gene has been inserted, recombinants...recombinants can first be selected via antibiotic resistance, but all clones should be further tested to confirm...
Showing: 201 - 220 of 234 results