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We narrowed to 507 results for: otos

Showing: 81 - 100 of 507 results

  1. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    Published
    April 20, 2021, 2:30 p.m.
    ...  Lowering cytotoxicity. Different envelope proteins have different levels of cytotoxicity and the mechanisms...pseudotype your viral vector, you use the same protocol for generating lentivirus, but you use a different...mechanisms behind the cytotoxicity can vary. For example, the popular VSV-G envelope protein causes cells... form syncytias that cause the cells to die. Cytotoxicity can also be cell type dependent: if a cell has... Pettie D, King NP, Balazs AB, Bloom JD (2020) Protocol and Reagents for Pseudotyping Lentiviral Particles...

  2. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    Published
    Nov. 5, 2014, 3:41 p.m.
    ...yield of the donor (number of emitted photons per absorbed photons) and the extinction coefficient of the...microscopy (FLIM) method. Acceptor photobleaching method The acceptor photobleaching method is simple but limited...commonly used to measure FRET: the acceptor photobleaching method and the fluorescence lifetime imaging... the donor is quenched when FRET occurs. By photobleaching the acceptor, you release the donor's quenching...

  3. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    Published
    Feb. 22, 2017, 3:30 p.m.
    ...reaction. Below is a step-by-step overview of the protocol: Prepare ssDNA template: The wild-type plasmid...depth. Can I use my favorite plasmid with this protocol? Maybe. Not all plasmids will work for the one-pot... BbvCI site(s). If so, the nicking mutagenesis protocol will still work if the orientation of the BbvCI...multi-site saturation mutagenesis with the same protocol. What do you think about one pot saturation mutagenesis...Methods in Molecular Biology In Vitro Mutagenesis Protocols, pg. 103-109. PubMed PMID: 20676978. 3. Wrenbeck...Scale Using Deep Mutational Scanning.” Nature Protocols 9.9 (2014): 2267–2284. PubMed PMID: 25167058. ...

  4. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    Published
    Jan. 29, 2019, 2:27 p.m.
    ... not photostable, and were presented in the study to support the uniqueness of the photostability of the... and non toxic to cells. Because it rapidly photobleaches, DFHBI can only be used for qualitative, but... Corn and its small molecule makes Corn more photostable than Spinach and Broccoli, and useful for quantitative...increased brightness compared to Mango, and all are photostable and useful for both live and fixed cell imaging...Imaging RNA polymerase III transcription using a photostable RNA–fluorophore complex." Nature chemical biology...

  5. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    Published
    May 5, 2020, 1:15 p.m.
    ...the target site, while Cas12a requires a PAM (Protospacer Adjacent Motif) for target cleavage. This test...40 min incubation. A scalable, easy-to-deploy, protocol for Cas13-based detection of SARS-CoV-2 genetic... Singh J, Miao X, Streithorst JA, Granados A, Sotomayor-Gonzalez A, Zorn K, Gopez A, Hsu E, Gu W, Miller... acid detection with CRISPR nucleases. Nature Protocols 14:2986–3012 . https://doi.org/10.1038/s41596-...Wilson, M. Z. (2020). A Scalable, Easy-to-Deploy, Protocol for Cas13-Based Detection of SARS-CoV-2 Genetic...Zhang, F., Abudayyeh, O. O. & Gootenberg, J. S. A protocol for detection of COVID-19 using CRISPR diagnostics...

  6. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    Published
    May 20, 2021, 1:15 p.m.
    ...production standpoint, it wasn’t ideal for our Lab Protocol videos. It’s really nice to have a scientist on-screen... on-screen, somebody who really knows the protocol and can guide viewers through. I know cameras, but ...thankfully volunteered, and we decided to focus on a protocol that didn’t require any lab shooting, just animation...setup for the lab, so we can get back to producing protocol videos?   Figure 4: The remote directing...the pandemic with some incredibly strict safety protocols to keep Addgenies safe. Space was at a premium...why not check out all of the written and video protocols at Addgene? If you need help in the lab, we have...

  7. In Living Color: The Skinny on In Vivo Imaging Tools

    Type
    Blog Post
    Published
    March 27, 2014, 5:14 p.m.
    ...last year, Verkhusha added two far-red light photoactivatable (PA) near-infrared fluorescent proteins (FPs...fluorescent proteins, iRFP670 and iRFP720, as photoacoustic contrast agents for two-color imaging in animals...genetically-encoded probes by making them brighter, more photostable, smaller, and less toxic to cells. They’d like...751-4. doi:10.1038/nmeth.2521) Far-red light photoactivatable near-infrared fluorescent proteins engineered...: 10.1038/ncomms3153. PubMed) Multicontrast photoacoustic in vivo imaging using near-infrared fluorescent...

  8. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    Published
    Jan. 24, 2019, 2:24 p.m.
    ...This post was contributed by Richi Sakaguchi from Kyoto University, and Marcus N. Leiwe and Takeshi Imai.... For more details on this procedure, find the protocol here.   Tips and troubleshooting your Tetbow...thanks to our guest bloggers Richi Sakaguchi from Kyoto University, and Marcus N. Leiwe and Takeshi Imai...University. Richi Sakaguchi is a graduate student at Kyoto University. He is interested in developing genetic...High-resolution fluorescence imaging using SeeDB2." Bio-Protocol 8 (2018): e3046. Livet, Jean, et al. "Transgenic...

  9. Fluorescent Protein Travel Awards - FLiPs and Fluorescent Protein Biosensors

    Type
    Blog Post
    Published
    July 27, 2017, 1:11 p.m.
    ...Xin Zhou greatly expanded that list by using photoswitchable fluorescent proteins to create enzymes that...turned the enzymes back on. Xin termed these photoswitchable enzymes Fluorescent Light-inducible Proteins...various designs, Xin has been able to develop photoswitchable Intersectin, Cdc42, Raf1, Mek1, Mek2, Cdk5,... You can learn more about Xin’s work with photoswitchable components of the ERK signaling pathway in ...Resources on the Addgene Blog Learn More about Photosactivatable Fluorescent Proteins Use Light to Control ...

  10. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    Published
    April 7, 2022, 1:15 p.m.
    ...sensor’s photophysical properties such as kinetics, ∆F/F0, SNR, dynamic range, photostability, quantum...signals upon sensing glutamate using both 1-photon and 2-photon imaging.   Figure 1: 1P and 2P excitation...membrane trafficking and localization. Improved photophysical properties The improved variants of iGluSnFR3...

  11. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

    Type
    Blog Post
    Published
    Nov. 6, 2018, 1:12 p.m.
    ...         Adipocytes [28] Keratinocytes [29] Photoreceptors [30] Otic hair cells [31] Neurons [32, 33... pluripotent stem cells from urine samples. Nat Protoc, 2012. 7(12): p. 2080-9. PubMed PMID: 23138349....from human ES and patient-specific iPS cells. Nat Protoc, 2015. 10(7): p. 941-58. PubMed PMID: 26042384....Generation, purification and transplantation of photoreceptors derived from human induced pluripotent stem...cerebral cortex neurons and neural networks. Nat Protoc, 2012. 7(10): p. 1836-46. PubMed PMID: 22976355...expression of insulin genes in liver and ameliorates streptozotocin-induced hyperglycemia. Nat Med, 2000. 6(5):...

  12. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

    Type
    Blog Post
    Published
    June 25, 2019, 12:54 p.m.
    ...experiment check out Addgene’s protocol page, which has both protocols and videos for techniques in basic...about making your own competent cells and our protocols page to learn about bacterial transformation in...plasmid cloning Resources on Addgene.org Read our protocols for plasmid cloning Find our molecular biology...biology reference here Find molecular biology protocols here ...

  13. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    Published
    March 23, 2021, 1:15 p.m.
    ...of Golden Gate cloning, the Haynes lab’s Protocols.io protocol describes an elegant system that allows ... Priode and Haynes, protocols.io. 2021. dx.doi.org/10.17504/protocols.io.brgcm3sw     New system developed...flanked by randomized nucleotides in place of a protospacer adjacent motif (PAM). They're used to determine...

  14. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    Published
    Feb. 4, 2020, 2:15 p.m.
    ... wavelengths of low photosynthetic activity. This still allows photosynthetically active wavelengths to... coral’s symbiotic dinoflagellates that are photosynthetic (Salih et al., 2000). Chromoproteins also enhance...Protein 101 blog series Learn more about FRET Find protocols and tips for making plasmid constructs Resources... (2000) Fluorescent pigments in corals are photoprotective. Nature 408:850–853 . https://doi.org/10.1038...

  15. Deep Dive: qPCR

    Type
    Blog Post
    Published
    Sept. 29, 2022, 1:15 p.m.
    ...included in commercial sets and most machines have a protocol that automatically performs the normalization ...normalization step.  Machines Most general-use protocols, unless otherwise specified, were likely developed on the...LightCycler, you’ll likely need to re-optimize protocols for that machine. (Teo, et al, 2002; see paper...an individually run sample when developing the protocol and optimized until the multiplexed and individual...samples or sample conditions change, the multiplexed protocol may have to be re-optimized.  While troublesome...eds.), Quantitative Real-Time PCR: Methods and Protocols, Methods in Molecular Biology, vol. 1160, DOI ...

  16. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    Published
    Oct. 18, 2018, 12:37 p.m.
    ...able to come of age. The in vivo assembly cloning protocol In Vivo Assembly (IVA) cloning (Garcia-Nafria ... to be performed using a simple two-step, 2 hr protocol prior to transformation (Figure 1). As this approach...procedures are performed using the same three step protocol:  Fragment generation - Any linear DNA fragments...plasmid cloning Resources on Addgene.org Read our protocols for plasmid cloning Find our molecular biology...biology reference here Find molecular biology protocols here  ...

  17. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    Published
    Aug. 30, 2022, 1:15 p.m.
    ...IF, look for general IF protocols for your sample type. Starting with a protocol that usually works in ...details to help you plan and optimize your own protocols. Here we’ll start with just the first couple of...cause some confusion. Paraformaldehyde: Many IF protocols call for paraformaldehyde (PFA) as the fixative...datasheet to see if it has been tested in IF and if a protocol is available. When possible, try to start with...

  18. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    Published
    Aug. 13, 2024, 1:15 p.m.
    ...Using a spectrophotometer is among the faster methods of quantification. Many spectrophotometers have an... Antibody Purification Protocol Addgene's Video Antibody Purification Protocol Addgene's Antibody Guide...Antibodies in Basic Neuroscience Research. Curr Protoc Neurosci. 2020 Dec;94(1):e106. doi: 10.1002/cpns...

  19. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ...Platinum Gate TALEN Kit, please see the Yamamoto lab’s protocol for TALEN construction. Sakuma et al...plasmids, watch informational videos, download protocols, and more! Looking for background information ...phosphoprotein system, which includes detailed protocols provided by the Rinehart lab. Steinfeld et al...the most recently deposited tools, resources, protocols, and more. MoClo modular cloning system Synthetic...assembly and activity? The laboratory of Takashi Yamamoto has created a complete TALEN assembly system after...overlap, minimal protein aggregation, and high photostabilty. When multiple cassettes are integrated, each...

  20. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    Published
    Aug. 14, 2018, 12:42 p.m.
    ...genetic tools to the brain because of its low cytotoxicity, low immunogenicity, and the long-term expression...The issue with these FPs is that they are less photostable than GFP. In addition, antibody-based amplification...fluorescence. Furthermore, many of these FPs are cytotoxic, prone to aggregation and cross-react with anti-GFP... CA3 pyramidal neurons which are known to be notoriously difficult to label. It has also been shown that...structure and color variants." Cold Spring Harbor Protocols 2009.12 (2009): pdb-top63. PubMed PMID: 20150100...
Showing: 81 - 100 of 507 results