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We narrowed to 472 results for: FLU;

Showing: 141 - 160 of 472 results

  1. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    ...Institute for Cell Science Plasmid Collection Fluorescent tagging strategy Figure 1: Gene editing... long stretches of DNA on either side of the fluorescent protein sequence that is homologous to the sequence...through homology directed repair To introduce the fluorescent tag to the cells, we used CRISPR/Cas9 to make...will attempt to repair itself without outside influence (non-homologous end joining, or NHEJ), in approximately...cells, and C. elegans. In addition to generating fluorescent protein fusions, plasmids are available for tagging...gene of interest. Endogenous gene tagging with fluorescent proteins as described here enables live cell ...about Non-Homologous End Joining Check Out Our Fluorescent Protein Posts Resources on Addgene.org Browse...

  2. Illuminating Epigenetics with A FRET Based Biosensor

    Type
    Blog Post
    ...either through histone modifications, which can influence the transcription of genes by RNA polymerase II...techniques include chromatin immunoprecipitation, fluorescent in-situ hybridization, methylation-sensitive ...facilitated by the flexible linker) that brings the fluorescent units of the FRET pair together and causes an...signal. Subsequent demethylation separates the fluorescent units and so lowers the FRET signal back again...Change in paternal grandmothers early food supply influenced cardiovascular mortality of the female grandchildren...Jao, and Alice Y. Ting. "Genetically encoded fluorescent reporters of histone methylation in living cells...the Ting Lab plasmids at Addgene Browse More Fluorescent Protein Biosensors Read Our Tips for Using FRET...

  3. Polymerase Chain Reaction Overview and Applications

    Type
    Blog Post
    ...probe based fluorescent dye that intercalates with any dsDNA and the use of a fluorometer feature built... thermocycler to measure that fluorescent output. With this fluorescent dye, the concentration of the ...which measures the fluorescence amplitude of each droplet. The fraction of fluorescent PCR-positive droplets...reaction cycles is continuously detected via a fluorescent signal. The signal increases proportionally to...concentration of the starting template DNA the fluorescent signal throughout the reaction is compared to...

  4. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    ...comparing the threshold fluorescence of the viral DNA to the threshold fluorescence of a standard - a set...plate and measures the fluorescence amplitude of each one. Droplets that fluoresce contain the amplified...been read, the software outputs an image of the fluorescence amplitude measured for each droplet in each ...droplet readers have a few channels for detecting fluorescence so that it is possible to measure the concentration...After additional rounds of amplification, your fluorescence amplitude will increase, leading to greater ...al. "A systematic investigation of parameters influencing droplet rain in the Listeria monocytogenes prfA...

  5. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...incubation, you assess the level of fluorescence in your cells under a fluorescence microscope. The higher the ... homology dependent repair by enhanced green fluorescent protein (EGFP) reconstitution. (a) Scheme of ...containing sgRNA sequences into HEK293T cells. The fluorescence intensity was classified into four groups (4...darker than control, 1; very dark). Brighter fluorescence indicates a more successful gRNA design. Adapted...your target is inserted, disrupting the EGFP fluorescent signal, for details on this plasmid, see Mashiko...the amount of fluorescence, the better that gRNA is at causing a double stranded break at the target site...screening as little as 8-10 pups. The level of fluorescence you saw in the validation assay should give ...

  6. Our Most Popular Blog Posts from 2019

    Type
    Blog Post
    ...CRISPR, optogenetics, chemogenetics, plasmids, fluorescent proteins, careers, and more. These resources ...Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins... tools used in cloning. 8. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi by Jennifer...and mWasabi and find plasmids containing these fluorescent proteins at Addgene. 7. Designing Your Chalk...chalk talk, and tips for practicing.  6. The Fluorescent Vegetables in Aptamer Soup by Aliyah Weinstein...aptamers, which are nucleic acid sequences that fluoresce upon binding a small molecule, are often named...making scientific graphics. PS: they’re free. 3. Fluorescent Proteins 101: GFP Fusion Proteins - Making the...

  7. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    ...when paired with appropriate fluorescent optics. Existing red fluorescent voltage indicators, however,...31937940. Detection of neural activity using a red fluorescent genetically encoded voltage indicator Article...voltage-sensitive inhibitory rhodopsin (Ace2N) with the red fluorescent protein (mScarlet) with either a seven or eight...manipulations when combined with existing green-fluorescent GEVIs and blue-light-sensitive channelrhodopsins...zebrabow. In this system, cells express various fluorescent proteins controlled by Cre recombination. Each...variety of colors depending on the combination of fluorescent proteins expressed. CreLite-mediated recombination...

  8. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

    Type
    Blog Post
    ...Resources Cre-lox Resource Guide Fluorescent Protein Resource Guide Fluorescent Protein Blog Posts ...constructs similarly to GFP, but commonly used red fluorescent proteins dsRed, mCherry, and TdTomato do not ...GFP and derivatives GFP and YFP, but not red fluorescent proteins, as seen with T-DDOGs. Like T-DDOGs...utliized as scaffolds, including the set of red fluorescent proteins. The T-DOGGs and Cre-DOG are available... C.Y., et al. “A nanobody-based system using fluorescent proteins as scaffolds for cell-specific gene ... Caussinus, E., Kanca, O., and Affolter, M. “Fluorescent fusion protein knockout mediated by anti-GFP ...

  9. A Primer on Optogenetics: Introduction and Opsin Delivery

    Type
    Blog Post
    ...co-localization immunofluorescence, cell-type specificity. Viruses that only express the fluorescent proteins...chloride ion pumps, activation of which results in efflux of chloride ions, subsequent hyperpolarization,...unlike HR, this is achieved through hydrogen ion efflux; evidence suggests that Arch may more effectively...Thankfully the Deisseroth constructs coexpress a fluorescent protein, which allows for easy identification...general rule of thumb, if you can detect a strong fluorescent signal (without antibody manipulation), then ... vectors for anterograde axonal tracing with fluorescent proteins in nontransgenic and cre driver mice...optogenetics plasmids! Find AAV resources! Find fluorescent protein resources!    ...

  10. Technical Design of a Western Blot

    Type
    Blog Post
    ...amount of protein you are expecting.  Fluorescent proteins Fluorescent proteins, while more expensive, allow...typically rely on primary antibodies conjugated to fluorescent proteins, instead of the more common chemiluminescent...you to optimize for smaller proteins or lower fluorescent backgrounds. However, PVDF does add a few (short... of reporters: chemiluminescence enzymes and fluorescent proteins. Chemiluminescence enzymes are cheap...important if you want to quantify your blots). Fluorescent proteins are stable, so blots can be stored for...magnitude, which includes a wide dynamic range for fluorescent proteins. They do tend to have increased background...usually more expensive Nitrocellulose or low-fluorescence PVDF membranes Increased fragility (nitrocellulose...

  11. Tag Your Favorite Yeast Genes with Ease

    Type
    Blog Post
    ...direct fusion of fluorescent proteins (FPs) to a yeast gene of interest. These fluorescently tagged genes ...several ready-to-use, modular plasmids, combining fluorescent tags, epitope tags, protease sites, and selection...of course—and start tagging! Yeast-optimized fluorophores for imaging Lee S, et al. (2013) PLoS ...marker. Lee et al.(1) assessed many of these fluorescent tags in S. cerevisiae, looking at their performance...requirements. Select from these yeast-optimized fluorophore tagging vectors for your single or multi-color...

  12. CRISPR Guide

    Type
    Collection
    ...catalytically inactive dCas9 to a fluorescent marker like green fluorescent protein (GFP), creating a customizable... sequences using a fluorescent reporter. In SHERLOCK, a quenched ssRNA fluorescent reporter is added to...: Epigenetics Visualizing Genomic Loci Using Fluorophores CRISPR imaging has numerous advantages, including...customizable DNA or RNA label for fluorescence microscopy. RNA aptamers, like MS2 or PP7, can also be fused...fused to the gRNA to recruit fluorescently-tagged RNA-binding proteins (RBPs). Multicolor CRISPR imaging...dCas9 and S. aureus dCas9) tagged with different fluorescent proteins or by fusing the gRNAs to orthogonal...imaging of genomic loci, as in the CRISPR-Sirius . Fluorescent CRISPR systems have been used for dynamic tracking...

  13. Using Ultrasound to Image Bacteria in vivo: Acoustic Reporter Genes

    Type
    Blog Post
    ... vivo imaging with fluorescent proteins Browse luciferase plasmids Find fluorescent protein biosensors...developed in vivo bacterial reporters that use fluorophores and luciferases to track bacteria in real time... reporters Limited Depth of Signal/Imaging: Fluorescent reporters are not as well suited for imaging ...context/spatial localization: In vivo imaging with fluorescent or bioluminescent reporters usually only detects...tools The luciferase reporter gene The green fluorescent protein reporter gene Resources on Addgene.org...

  14. Understanding GPCR Signaling Using TRUPATH

    Type
    Blog Post
    ...FRET), in which a donor fluorophore is used to excite an acceptor fluorophore. The difference for BRET...blog Browse all of our fluorescent protein blog posts Find other fluorescent biosensors Resources on...transfer is between a luminescent donor and a fluorescent acceptor. TRUPATH uses an updated version of ...on Addgene.org Browse the catalog of fluorescent protein plasmids Find biosensors for your experiments...

  15. CRISPR Plasmids for Genomic Visualization

    Type
    Collection
    ...catalytically inactive Cas9 (dCas9) fused to a fluorescent marker like GFP, researchers have turned dCas9...into a customizable DNA labeler compatible with fluorescence microscopy in living cells. Alternatively, gRNAs...specific RNA-binding proteins (RBPs) tagged with fluorescent proteins to visualize targeted genomic loci. ...live cell imaging. Compared to techniques like fluorescence in situ hybridization (FISH), CRISPR imaging... the chromatin dynamics in living cells. The fluorescent CRISPR system has been used for dynamic tracking...

  16. RANbodies: Reporter Nanobody Fusions

    Type
    Blog Post
    ...and instead are conjugated to reporters (i.e. fluorescent dyes, HRP, etc. ) after purification. Antibodies...signal amplification occurs when HRP converts a fluorescently labeled tyramide molecule into a highly reactive...linked to residues within close proximity thus fluorescently labeling them. HRP RANbody staining results ... ≥100-fold increase in signal over intrinsic fluorescence. Increased signal is likely due to lower background...and tissue sections Amplifying weak signals of fluorescent protein (i.e. GFP, RFP) Detecting GFP reconstitution... from Penn State Need help picking the right fluorescence microscopy technique to use? Check out this ...

  17. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    ... in bacteria CRISPR-Casɸ mGold, a new yellow fluorescent protein New CRISPR plasmids New items from the...32675376/ mGold, the most photostable yellow fluorescent protein to date By Justin Ng High-throughput ...High-throughput screening methods, such as fluorescence-activated cell sorting (FACS) helps researchers sift through...million cell types expressing variants of Yellow Fluorescent Protein (YFP). The team identified a new YFP ...Resources page.  CDDR (CRISPR–Cas9-based Dual-fluorescent DSB Repair) is a double-strand break repair assay...the introduction & resolution of two DSBs in a fluorescent reporter. Find the plasmids here Yongsub Kim...

  18. Recombinases AAV Preps

    Type
    Collection
    ...these molecular tools. Cre AAV ID Name Promoter Fluorophore/Tag Serotype(s) PI 105551 pENN.AAV.CamKII.HI....AAV.TBG.PI.Cre.rBGe TBG none 8 Wilson Dre AAV ID Name Promoter Fluorophore Serotype(s) PI 50363 AAV phSyn1(S)-DreO-bGHpA...Syn none 5, rg* Zeng Flpo AAV ID Name Promoter Fluorophore Serotype(s) PI 51669 AAV phSyn1(S)-FlpO-bGHpA...Light-Inducible Recombinases ID Name Promoter Fluorophore Serotype(s) PI 140135 pAAV-EF1a-iCreV EF1a none...none 1, PHPeB Zeng VCre AAV ID Name Promoter Fluorophore/Tag Serotype(s) PI 55638 pAAV-EF1a-vCre EF1a ...

  19. Hot Plasmids: Fall 2024

    Type
    Blog Post
    ... highly autofluorescent tissues like the intestine. Figure 3: Expression of fluorophore-tagged ... imaging in C. elegans By Alyssa Neuhaus Red fluorescent proteins like mScarlet are essential tools for...2024 under CC-BY license.  These optimized fluorescent proteins will be a useful addition to the toolkit...Comparative analysis of new mScarlet-based red fluorescent tags in Caenorhabditis elegans. Genetics, 228...wavelength (460 nm) is shorter than many popular fluorescence markers like GFP (510 nm), allowing co-imaging...

  20. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    ...quenched fluorescent ssRNA reporter is added to the reaction. Cleavage of the quenchable fluorescent RNA by...Cas13 and Cas12a, are combined with different fluorescent reporters for each enzyme. In addition, Csm6,...Csm6 and a Csm6 specific reporter (in the same fluorescent channel as the Cas13 reporter), they could significantly...papilloma virus (HPV) genome, via a crRNA. An ssDNA-fluorescently quenched (FQ) reporter, which will produce a...rate of ~1,250 cuts per second. A quantifiable fluorescent signal designates the presence of your DNA of...
Showing: 141 - 160 of 472 results