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We narrowed to 379 results for: Lor

Showing: 31 - 60 of 379 results

  1. Kit Free RNA Extraction

    Type
    Protocol
    ...are also included: using Isopropanol or Lithium Chloride. This protocol was adapted from (Link opens in...Water-saturated Phenol 2 M Sodium Acetate pH 4 Chloroform/Isoamyl alcohol (49:1) 75% Ethanol RNase-free...for precipitation step, Option A) 7.5 M Lithium Chloride (for precipitation step, Option B) Glycogen (Optional...phenol, mix thoroughly by inversion. Add 0.2 mL of Chloroform/Isoamyl alcohol (49:1) and then shake vigorously...(s). You can use either Isopropanol or Lithium Chloride for this step. Isopropanol (Option A): Add 1 volume...aqueous layer. Incubate at -20°C for 1 hour. Lithium Chloride (Option B): LiCl selectively precipitates RNA ...from the homogenized sample(s). Add 0.2 mL of Chloroform/Isoamyl alcohol (49:1) per 1 mL of TRIzol® used...

  2. Lentivirus Production

    Type
    Protocol
    ... add hydrochloric acid until the solution clears. Check the pH of the solution. Use hydrochloric acid ... Opti-Pro SFM, Thermo Fisher, 12309019 25 mM chloroquine Polyethylenimine, linear MW 25,000 Da Heat-inactivated...Corning 430167 15 mL conical tubes, VWR 21008-216 Hydrochloric acid Sodium hydroxide 0.22 μm polyethersulfone... will be basic and will need adjustment with hydrochloric acid first. Pro-Tip The pH of this solution ...mL conical and an appropriate amount of 25 mM chloroquine to the conical so that the final concentration...transfection mixes are added) will be 25 uM of chloroquine. After the incubation, add the DNA:PEI-Max mix...mix to the conical containing the media and chloroquine and mix well. Gently aspirate the media out of...

  4. Cre-lox system

    Type
    Collection
    ...marking of mouse neurons with multiple distinct colors Mammalian Sanes See article Multibow constructs...dependent tdtomato expression. See article for more colors. Mammalian Zeng 24334 hsp70l-loxP-mCherry-STOP-...

  5. Pouring LB Agar Plates

    Type
    Protocol
    ...Pro-Tip We batch label our plates with colored marker - particular colors correspond to particular antibiotics...extract 10.0 g peptone from casein 10.0 g sodium chloride 12.0 g agar-agar 1 L Sterile H 2 O Sterile plates...5 µg/mL Carbenicillin* 100 mg/mL 100 µg/mL Chloramphenicol 25 mg/mL (dissolve in EtOH) 25 µg/mL Coumermycin...

  6. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...periodically to determine when the desired color change has occurred. The color will change from yellow to blue ...optimize experimental conditions. When the desired color change has occurred, gently remove the plate seal...generate a standard curve . If you don’t see any color change or dose response, your antibody concentration...

  7. AAV Production in HEK293 Cells

    Type
    Protocol
    ...add hydrochloric acid until the solution clears. Check the pH of the solution and use hydrochloric acid... pH 8.5 + 60 mL of 5 M Sodium Chloride + 4 mL of 1 M Magnesium Chloride Close the bottle and mix by inverting...

  8. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    ...CRISPR to label multiple loci in distinct colors. To create a colorful Cas9 toolbox, Ma et al. turned to SpCas9...nucleome. CRISPRainbow: Scaling up to 6 colors + white Color Hairpin Blue MS2-MS2 Green PP7-PP7 ...number of limitations. Since color is specified by PAM sequence, each color requires a different Cas9 ortholog...Colorful CRISPR technologies are helping researchers visualize the genome and its organization within...track multiple genomic loci over time using all the colors of the CRISPRainbow. Figure 1: CRISPRainbow...chromosome-specific loci in the U20S cancer cell line./ Each color represents a gRNA targeting a specific chromosome...different fluorescent protein to create three distinct colors. The specificity of these orthologs is key: since...

  10. Antibodies 101: Flow Cytometry Controls

    Type
    Blog Post
    ...later. Single Colors Single color controls contain, as the name suggests, a single color from your antibody... single color, fluorescence minus one (FMO), and isotype. You should always have single color controls...single color control for every fluorophore you intend to use in your panel. Using single color controls...from your single color controls may not be accurate for your samples. The single color controls should ... flow plots for a single color control. A) A flow plot showing a single color control for the APC channel...can bind to any primary antibody you add. Single color controls are used to help you set up your voltages...achieved by adjusting your voltage using your single color controls (Figure 1A), instead of using your actual...

  11. AAVs CREATed for Gene Delivery to the CNS and PNS

    Type
    Blog Post
    ...diversity of colors AND sparse labeling of neurons , Chan et al separated color labeling and color expression...these AAV’s be used for?  1. Multi-color labeling of neurons Multi-color labeling of neurons is for more ... high levels of color diversity so that cells are labeled with their own unique color; and 2) low levels...labeling, without loss of color diversity.  With this two part system, color diversity is maintained at... neurons. A simple but flawed approach to multi-color labeling is to inject a mix of AAV vectors that ...multiple viral particles and labeled a variety of colors (figure 4b). But notice in figure 4b how difficult...AAV and red, green, or blue are the predominate colors expressed (figure 4c).   AAV Vector  Percent...

  12. Institute for Protein Innovation

    Type
    Collection
    ...anti-glypicans that accurately detect glypican presence and explore glypican function, providing a reliable and accessible...

  13. Optogenetics Plasmid Collection

    Type
    Collection
    ... cells. Other microbial opsins, such as the Halorhodopsins, Archaerhodopsins and Leptosphaeria rhodopsins...

  14. Affinity Purification of Recombinant Antibodies with Protein A or Protein G

    Type
    Protocol
    ...dibasic, Millipore Sigma S7907-500G 1 M Trizma hydrochloride solution pH 9.0, Millipore Sigma T2819-1L 250...binding buffer 2x . Add 50 µL of 1 M Trizma hydrochloride pH 9.0 to 10 microcentrifuge tubes. Cap the ... into the tubes containing 50 µL 1 M Trizma hydrochloride pH 9.0. Cap the tubes and vortex for 5 s to ...

  15. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    ... decided to jump into one of those unexplored areas with our color-changing flower, but to do that we’...plants can be challenging, there are a lot of unexplored areas in plant biotechnology that are ripe with...specifically for consumer sales. Flowers that change color throughout the day cannot be developed through conventional...years of basic research standing behind it. Our color changing flower involves two basic components: the...these components to develop flowers that change color. Without this basic research – this comprehensive...have also been patented. In order to construct our color changing flower we are carefully making our way ...Revolution Bio had to address as we developed the color-changing flower, and we’d love to hear about the...

  16. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ... to Image>Color>Replace Red with Magenta. Or you can use Color>Split Channels and then Color>Merge Channels... are accessible to all viewers. Use a colorblind-friendly color palette for display (like green-magenta-blue...secondary antibodies, and image all the different colors in one session – fast and easy! Just make sure ...isotypes and that each secondary has a different color fluorophore that you can image separately on your... the images of each labeled target as different color channels and analyze the results. Depending on the...show each channel in individual panels, then the multicolor overlay; this helps all viewers, especially if...automatically from the microscope software, you can always recolor them! In Fiji, for example, you’d open the offending...

  17. A Guide to Designing a Scientific Poster: Content, Layout, and More

    Type
    Blog Post
    ...and color One of the best ways to grab your audience’s attention is with the background and color of your...Depending on the color you select for the background, you’ll want to make sure that the color of your font...background, but two common approaches are either a solid color, or a “themed” background that is relevant to the...but beware. A distracting background or clashing colors can take away from what should be the focus of ...the data and your story. If you choose a single-color background but want to add a bit more “flash,” you... in all of our illustrations above) so that the color fades to a lighter or darker version of itself from... is compatible. Use high-contrast between these colors so that your text is easily read. Similarly, be...

  19. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    ...background on the color spectrum and the principles of excitation and emission. The visible color spectrum is...principle of color emission. As you can see in Figure 1, emission spectra of neighboring colors can and do... Figure 1: Emission spectra and respective colors of a few commonly used fluorophores. Each fluorophore...flow-cytometry-fluochrome-brightness-spillover Excitation Laser Color Very Bright Bright Moderate Dim Ultraviolet...fluorophore-conjugated antibodies together in the same color panel. In this case, make sure to compare the excitation... issue. Make sure to check out our blog post on color compensation to learn more about them. Alright, ... dead cells present and it is fine to use a dim color, like APC-Cy7. Next comes our transduction reporter...

  20. Visualizing Genomic Loci with CRISPR-Sirius

    Type
    Blog Post
    ...Massachusetts Medical School developed a dCas9-based multicolor labeling system to visualize chromosomal loci...two different hairpins will produce a secondary color (yellow, cyan, or magenta) depending on the hairpin... containing all three hairpins produces a white color. While CRISPRainbow allows for simultaneous visualization...inserted into the sgRNA generates blue, green, or red colors for labeling. A combination of MS2, PP7, or boxB...sequences in the same sgRNA will generate additional colors. Image from Ma et al., 2016 with permission. ...fluorescent ligands to the loci of interest. Dual color detection In order to label two loci simultaneously...CRISPRainbow) to 3-fold with CRISPR-Sirius. Triple color detection With a third sgRNA containing alternating...

  21. CRISPR Plasmids - Prime Edit

    Type
    Collection
    ... improve the efficiency of editing, including exploring new sources for the reverse transcriptase. Many...

  22. RNA Extraction Without A Kit

    Type
    Blog Post
    ... using cesium-chloride gradients, water-saturated phenol, sodium acetate, and chloroform are added to ...material. Then, the homogenate was loaded onto cesium-chloride gradients and ultracentrifuged for up to 20 hours... protocol with guanidinium thiocyanate-phenol-chloroform. In this method, samples are still homogenized...ultracentrifugation!), the layers of phenol and chloroform separate, and RNA is retained in the top, aqueous...centrifugation and the supernatant is extracted with chloroform as in the above no-kit method.  Researchers have... trick is to precipitate the RNA using lithium chloride. LiCl solutions selectively precipitate RNA, but...Isolation by Acid Guanidinium Thiocyanate–Phenol–Chloroform Extraction. Analytical Biochemistry 162:156–159...

  23. Antibodies 101: ELISA (Enzyme-linked Immunosorbent Assay)

    Type
    Blog Post
    ...added to the wells. This reaction produces a colorimetric reaction that reflects the concentration of .... The reaction is stopped by the addition of hydrochloric acid (for more details on this, see below). ... concentration or antibody specificity via a colorimetric reaction. Image from Boguszewska et al., 2019... concentration or antibody specificity via a colorimetric reaction. Image from Boguszewska et al., 2019...added and the reaction can be visualized via a colorimetric reaction. The direct ELISA is often used when...

  24. Story of a SynBio Startup: RevBio's Epiphany (or Lack Thereof)

    Type
    Blog Post
    ...no one has ever seen before (flowers that change color throughout the day) and doing this with very little.... Our scientific training was put to use as we explored ideas, planned out potential strategies for pursuing...Anything was fair game for speculation - in fact, our color changing flower project was inspired by a Facebook...upon the idea of building a flower that changed color throughout the day. It would be a beautiful GMO,...we didn’t figure out how we would accomplish the color change until a few months later. RevBio accelerates...clue how to run a business. SynBio Axlr8r thought color-changing flowers was a great business idea as well...the funds needed to build a flower that changes color, but the experience is invaluable. This opportunity...

  25. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    ...fluorescence states, they can change from one fluorescent color to another, or their fluorescence can be switched...PC-FPs). PC-FPs can convert from one fluorescent color to another (e.g. green to red, or cyan to green)...Protein Oligomeric State Activating Light Pre/ Post Color Contrast (Fold Change in Fluorescence) Brightness... a red fluorescent state make them useful for 2-color imaging with green PA-FPs. Table 2: Properties of...experiment even begins! If you are performing multicolor experiments, you will want to be sure that the...Photoactivatable mCherry for high-resolution two-color fluorescence microscopy." Nature methods 6.2 (2009...

  26. CRISPR Plasmids - Bacteria

    Type
    Collection
    ...Marraffini 42876 pCas9 BsaI E. coli, S. pneumoniae Chloramphenicol yes, cut Marraffini 44251 pgRNA-bacteria BBa_J23119...

  27. Fluorescent Protein Guide: FRET

    Type
    Collection
    ...interest and the listed fluorescent protein. Plasmid Color Expression Description pPROEX Aqua Cyan Bacterial...

  28. Mapping the 4D nucleome with CRISPR/Cas9

    Type
    Blog Post
    ...limitation of this system is the single-color labeling; multicolor labeling would enable the visualization...every week! One of the newest applications is a colorful system that uses fluorescently labeled Cas9 to...may be altered in disease states. dCas9 gets a colorful makeover Even without its nuclease activity, catalytically...the cell cycle or other stimuli.   To create a colorful Cas9 toolbox, Ma et al. turned to SpCas9 and its...a different fluorescent protein to create three colors. The specificity of these orthologs is key: due...excited to see what’s coming next! References Multicolor CRISPR labeling of chromosomal loci in human ...

  29. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ...module_attribute "style" is_json="true" %}{% raw %}{"button_color":{"color":"#0052a9","css":"#0052a9","hex":"#0052a9...size_unit":"px","css":"color: #FFFFFF;font-size: 19px;font-family: helvetica;","color":"#FFFFFF","variant...text-decoration: none","font_set":"DEFAULT"},"button_hover_color":{"color":"#0052a9","css":"rgba(0, 82, 169, 75%....75)"},"override_default_style":true,"text_hover_color":{"color":"#ffffff","css":"#ffffff","hex":"#ffffff...

  30. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...Glutamate indicators with increased sensitivity and tailored deactivation rates Addgene's own empty lentiviral...Glutamate indicators with increased sensitivity and tailored deactivation rates By Mike LacyGlutamate is one...Reporter (iGluSnFR) designed and improved by the Loren Looger and Kaspar Podgorski Labs now has a new version...Glutamate indicators with increased sensitivity and tailored deactivation rates. bioRxiv, 2025.03.20.643984.../10.1101/2025.03.20.643984.  Build your way: Explore Addgene's new empty backbones By Meghan Rego You...cell line creation (Figure 2). Looking for more? Explore our empty and EGFP retroviral plasmid set, our ...(TMT) mass spectrometry (MS) in HCT116 cells (colorectal cancer). The bottom panel depicts the identification...
Showing: 31 - 60 of 379 results