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We narrowed to 729 results for: NSI

Showing: 81 - 90 of 729 results

  1. Plasmids 101: Mammalian Vectors

    Type
    Blog Post
    Published
    March 25, 2014, 3:15 p.m.
    ...resultant cells can either express the plasmid DNA transiently (similar to bacteria) or incorporate the genetic...ever-versatile fluorophore in a later post, so stay tuned! Transient transfection and the elusive "mammalian ORI" For... for the transfected plasmid to be expressed transiently. Since the DNA introduced in the transfection...want the plasmid depleted, but still want to use transient transfection methods. Since there are no "natural... generations. Stably transfected cells may be considered an entirely new cell line from that of the original...plasmid contains a cassette that will make cells sensitive to a toxin). In the table below we focus on positive...causes strand scissions 100-400 ug/mL *Not comprehensive.   ** In eukaryotes.   ***The concentration ...

  2. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    Published
    April 16, 2024, 1:15 p.m.
    ...GCE4All: Making Genetic Code Expansion Accessible Addgene's Genetic Code Expansion Collection...protein tags. The basics These tags most often consist of an enzyme that has been mutated to block its...CP appearance at the filament tip from (a). c) Intensity traces at the tip of the filament from (a-b), ... effects on actin filament length (lower). d) Intensity and filament length traces for a different event...of organic dyes or specialized ligands is more expensive than letting the cells make a fluorescent protein...Covalent Labeling of Recombinant Protein Molecules Inside Live Cells. Science, 281(5374), 269–272. https:... W. (2011). Chemical Tags for Labeling Proteins Inside Living Cells. Accounts of Chemical Research, 44...

  3. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    Published
    March 6, 2018, 1:49 p.m.
    ...or qPCR-based methods to greatly increase the sensitivity of detection (14-20). LDR has also been extended... do not read specific base sequences, but are sensitive to distortions in helix shape (22). Large purine... can be achieved through a combination of the intrinsic selectivity of the ligase active site and careful... that extends earlier studies through a high-sensitivity multiplexed format (21, 27). This methodology...In this method, substrate pools were prepared consisting of one target (template) strand and four upstream...Eukaryotic DNA ligases: structural and functional insights." Annu. Rev. Biochem. 77 (2008): 313-338. PubMed...JS, et al. "A high-throughput assay for the comprehensive profiling of DNA ligase fidelity." Nucleic acids...

  4. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    Published
    March 7, 2017, 3:30 p.m.
    ... cells are sensitive, it might be more difficult to get them to grow out. The only downside is that it.... In theory you could use densitometric analysis to quantitate the intensity of the bands on your gel ...reagent-grade endangered Martian chicory root, but considering this is a needle in a haystack type of objective...In retrospect I used a way too high Cas9 MOI considering that A549 cells are relatively easy to transduce...stable cell pool (pool) and from individual cell expansions (monoclonal, 1-5). Wild-type A549 cells do not...followed. If you have never used this assay before, consider following their tips, including: Before digesting...

  5. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

    Type
    Blog Post
    Published
    March 19, 2019, 1:31 p.m.
    ...turned to optogenetics. Through the use of light-sensitive protein domains, they have developed a system .... They accomplished this by inserting a photo-sensitive LOV2 domain from A. sativa into a loop region ...brain. But this process is time consuming and expensive. Thus scientists have turned to genetically encoded...electrical activity via imaging. They took the light-sensitive protein QuasAr2 and systematically mutated it....localization, has high signal:noise ratio, high sensitivity, responds quickly, is photostable, and is compatible...interaction (PPI) studies in plants that rely on transient transfection systems using Agrobacterium (such...Grefen and Blatt, 2012). The 2in1 Plasmid Toolkit consists of 4 plasmid toolkits generated with 4 different...

  6. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

    Type
    Blog Post
    Published
    Aug. 5, 2021, 12:15 p.m.
    ...recently developed a third-generation intensiometric glutamate sensing fluorescent reporter (iGluSnFR3) to...and their research.   Abhi Aggarwal: Glutamate sensing reporters Abhi Aggarwal is a postbaccalaureate ...new version of the biosensor is faster and more sensitive than its predecessors. Currently, Aggarwal, and...Research Center (DKFZ) who has developed new redox sensitive GFP2 (roGFP2)-based biosensors for live cells.... to better understand the processes that happen inside the cell,” says Pedre Pérez. Figure 2: ...changes following opioid withdrawal, providing insight into new methods for treating patients in recovery...

  7. A Primer on Optogenetics: Introduction and Opsin Delivery

    Type
    Blog Post
    Published
    Sept. 10, 2015, 2:30 p.m.
    ...methodological and experimental limitations and considerations? I recently spent two months in the lab of ... at NIDA and some of the practicalities and considerations you may not find in the materials and methods...use optogenetics The most important question to consider is what exactly do you want to use optogenetics...are also available. Picking your opsin  You must consider whether you wish to stimulate or inhibit neuronal...include Channelrhodopsin2 (Chr2), a blue-light sensitive non-specific cation channel, activation of which...potential. Halorhodopsins (HR) are green/yellow-light sensitive chloride ion pumps, activation of which results...breeding, and maintenance of a mouse line is also an expensive and time-consuming endeavor. Finally, expression...

  8. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    Published
    June 27, 2017, 1:03 p.m.
    ...within 10 nanometers of each other, or measure the tension in the helical structure of spider silk, or the...As the choice of fluorophore pairs is a key consideration for a FRET experiment, the possibilities can...Förster resonance energy transfer by monitoring sensitized emission in living plant cells." (2013). PubMed...linker sequence between the donor and acceptor is sensitive to a change in the environment which alters the...processes and signaling cascades, quantify mechanical tension (a molecular “spring”) (Meng et al., 2008), and...animals (Hirata and Kiyokawa 2016), and molecular tension microscopy (MTM), which could monitor physical ...Charlène, and Borghi, Nicolas. "FRET-based molecular tension microscopy." Methods 94 (2016): 33-42. PubMed PMID...

  9. The Materials Science of Optogenetics Experiments

    Type
    Blog Post
    Published
    Sept. 17, 2015, 2:30 p.m.
    ...for optogenetics. There are several important considerations with fabrication of the fiber optics needed...stimulation. The scatter effect is also an important consideration when preparing the implant itself, which brings...will be erratic, opsin activation/inactivation inconsistent, and behavioral results impossible to interpret...sequentially increasing fineness helps to maintain a consistent stream of light. Test your implant by connecting...least 85% of the initial laser output. The same considerations apply to the polishing and preparation of the...: closed/light receptive, open, closed/light insensitive. If you deliver a continuous stream of photons...optogenetics we’ll delve into some of the important considerations when performing optogenetics behavioral experiments...

  10. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    Published
    June 22, 2017, 2:30 p.m.
    ...structures, called organelles. Take a sneak peak inside a cell with the images from the Allen Cell Explorer...and diseased states can also provide interesting insights into disease mechanisms and protein function. ...complexes with. These “colocalization” studies give insights about the proximity of two proteins within the...cameras and image analysis software are highly sensitive to detecting light signals not visible to the ...human eye and are indifferent to subjective color intensity perception. Open source image analysis software... and well-executed experiments require some consideration of FP properties. It is important to look out...PMC4777285. Further Reading Microscopy U. Generfal Considerations when Using Fluorescent Proteins. Ruusuvuori...
Showing: 81 - 90 of 729 results