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We narrowed to 771 results for: Dos

Showing: 701 - 720 of 771 results

  2. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ...Gently mix sample (do not vortex) Incubate 30 min at 37 °C Transfer to ice ** Critical: do NOT treat your ...plates) and store at -20 °C. Once a standard is thawed do not freeze it again but store at 4 °C and use within... duplicate Load 5 μL of each sample in duplicate. Do not forget to include a no template control ( NTC...

  3. Protocol - How to Streak a Plate

    Type
    Protocol
    ...broad stroke. Only touch the surface of the plate, do NOT dig into the agar. Another very popular technique...bacterium. If the bacterial growth is too dense and you do not see single colonies, re-streak onto a new agar...

  4. Exploring Applications of the Bioluminescent HiBiT Tag

    Type
    Blog Post
    ...surface epitopes for many proteins, but some proteins do not have such epitopes. These classical 'undruggable... to track endogenous activity of EGFR, including dose-dependent response to EGF (Promega, 2025). The value...diminished luminescence signal, associated with endosomal localization of EGFR. This method offered kinetic...

  6. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ...systems listed above are orthogonal, meaning that they do not affect each other. For example, the GAL4 transcription...

  7. Protocol - How to Perform Sequence Analysis

    Type
    Protocol
    ...tags, mutations and a portion of the insert, but we do not sequence the entire plasmid. Addgene strongly... match Addgene’s sequencing result, what should I do? Check your trace file first; the apparent mismatch...

  8. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ... disperse mixture evenly. Do not pipette or swirl too vigorously, as you do not want to dislodge the cells...antisense sequences from step B.1 into the oligos below. Do not change the ends; these bases are important for...12-15 hours. c. In polypropylene microfuge tubes (do NOT use polystyrene tubes), make a cocktail for each...first. Pipette FuGENE® directly into the OPTI-MEM – do not allow FuGENE® to come in contact with the walls...media through a 0.45 μm filter to remove the cells. Do not use a 0.2 μm filter, as this is likely to shear...virus-containing media and replace with fresh media. Do not add puromycin until at least 24 hours after infection...

  9. AAV Production in HEK293 Cells

    Type
    Protocol
    ...yield. Do not overgrow your cells. Pass the cells twice a week during the maintenance phase and do not allow...Rapid-Flow PES Filtration Unit, Nalgene 167-0045) Pro-Tip Do not use filters made of materials other than PES..... AAV particles stick to many other surfaces, but do not stick to PES. Using a PES filter will maximize...

  14. Pouring LB Agar Plates

    Type
    Protocol
    ...bacteria containing that plasmid from bacteria that do not contain it by artificial selection (i.e. growing... of the bottle with its cap or aluminum foil (but do not make an air-tight seal!) and tape the bottle ...molten gel-mix in the water bath for at least 5 min. Do not let any of the water bath water touch the neck...

  15. Fluorescence Titering Assay

    Type
    Protocol
    ...regularly Do not over- or under-grow your cells. Thaw a new vial of cells after 20–30 passages. Do not add...

  16. Protocol - Bacterial Transformation

    Type
    Protocol
    ...transforming large plasmids (>10 kb) or BACs, what can I do? Chemically competent cells are fast and easy to ...cell/DNA mixture to induce membrane permeability. To do this you will need to have access to an electroporator...

  17. Which Fluorescence Microscopy Technique is Best for Me?

    Type
    Blog Post
    ...μm) tissue sections and 3D cultures These samples do not necessarily require optical sectioning, but open...while also avoiding the toxic effects of high light doses to the cells. Thin static samples Ex: Fixed monolayers...out of focus light. However, they also continually dose the sample from top to bottom with excitation light...

  18. Pipetting Protocol

    Type
    Protocol
    ...box and push the end of the pipette onto the tip. Do not touch the tips with your fingers to avoid contaminating...plunger, gently lower the pipette tip into the liquid. Do not submerge the pipette itself into the liquid and...

  19. Colony Formation Titering Assay

    Type
    Protocol
    ...regularly Do not over or under-grow your cells. Thaw a new vial of cells after 20–30 passages. Do not add...Procedure Before beginning a colony formation assay, the dose of antibiotic required to kill your target cell ...determined. Treat the target cells with a range of doses of antibiotic. Determine the minimum concentration...window) required to kill all of the cells. Use this dose for the colony formation assay. Prepare a batch ...

  20. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...thicker as the proteins and DNA are denatured. Pro-Tip Do not vortex at this stage or the genomic DNA will ...4. Note: Phenol-chloroform is a hazardous waste - DO NOT pour down sink. Top Protocol: Ethanol Precipitation...
Showing: 701 - 720 of 771 results