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Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

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We narrowed to 916 results for: IRE;

Showing: 861 - 880 of 916 results

  13. Lab Safety for Biosafety Levels One and Two

    Type
    Protocol
    ...Sharps container Biohazard waste container Fire blanket Fire extinguisher Guidelines BSL-1 Guidelines Before...eyewash station, safety shower, fire blanket, and extinguisher are also required to be present in the room....each level, as each level has different safety requirements. BSL-1 is designated for those working with ...times when you may be working with a protocol that requires shaking or mixing, which may produce aerosols ... PPE protocols . Working in a BSL-2 laboratory requires laboratory glasses in addition to coat and gloves...room and on the door. BSL-2 centrifugation steps require an aerosol-tight lid and rotors should be loaded...in a biosafety cabinet. BSL-2 safety protocols require bloodborne pathogens training. It is strongly recommended...

  14. Western Blot

    Type
    Protocol
    ...stringent buffers and a sonication step may be required for hard to extract proteins such as those in ...standard curve. Determine the volume of sample required to load an equivalent amount of total protein ...continue running the gel until you have obtained the desired separation. Gently open the gel with the spatula...Device. Close the lid of the device. Select the desired method and make sure the parameters are correct... incubation Dilute the primary antibody to the desired concentration in blocking buffer. Pro-Tip The optimal...peroxidase-conjugated secondary antibody to the desired concentration in blocking buffer. Pro-Tip The optimal...

  15. Kit Free RNA Extraction

    Type
    Protocol
    ... as bone, or bacteria/yeast/plant samples will require additional steps to effectively lyse open the cells...wash and add RNase-free water or TE as soon as the entire tube is dried but while the white pellet is still...Pro-Tip To avoid multiple freeze-thaw cycles of your entire RNA sample, consider making smaller aliquots of...

  16. CRISPR Library Amplification

    Type
    Protocol
    ...possible but note that individual libraries may require modifications dictated by the originating laboratory... can be scaled according to the actual number. Required Quality Control (QC) Pooled libraries can be challenging...a smaller plasmid containing only the elements required for bacterial propagation (origin of replication...100,000 plasmids. Colonies may appear small and require extra incubation time in order to be enumerated...colonies but as long as one can ensure more than the required colonies are present. If colonies are not present...can dramatically plateau and sometimes fall off entirely if the column or reagents are significantly overloaded...

  17. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ... RNAi Consortium has built a library of shRNAs directed against 15,000 human and 15,000 mouse genes. Addgene...shRNAs. pLKO.1 can be introduced into cells via direct transfection, or can be converted into lentiviral...protein expression, you may transfect plasmid DNA directly into the target cells. The shRNA will be expressed...OPTI-MEM necessary given that each reaction will require 6 μL FuGENE® + 74 μL OPTI-MEM. For example: 1x ...polypropylene tube, add OPTI-MEM first. Pipette FuGENE® directly into the OPTI-MEM – do not allow FuGENE® to come...for a total volume of 100 μL. Pipette master mix directly into the liquid and not onto the walls of the ...

  18. Protocol - How to Perform Sequence Analysis

    Type
    Protocol
    ...verify the ends of the insert. Longer genes may require internal primers that anneal to a specific region...portion of the insert, but we do not sequence the entire plasmid. Addgene strongly recommends that you...Make sure the primer only anneals once within the entire construct. Is the primer an appropriate distance... within the Addgene sequencing results? If you require additional sequencing and need to design a custom...

  19. Protocol - How to Inoculate a Bacterial Culture

    Type
    Protocol
    ...bottle may explode!) and then loosely cover the entire top of the bottle with aluminum foil. Autoclave...shaking incubator. Note: Some plasmids or strains require growth at 30°C. If so, you will likely need to ...in the media (see right). Notes: Some protocols require bacteria to be in the log phase of growth. Check...bacteria incubated at 30°C rather than 37°C often require longer incubation times. Double check that the ...

  20. Centrifugation

    Type
    Protocol
    ... in 1.5 mL microfuge tubes at 10,000 rpm would require a tabletop microcentrifuge like the one in the ... you are working. Even if your samples may not require specific PPE, you never know what else has been...RPM), these are not the same units and are not directly interchangeable. If you need to convert between...supernatant. Depending on your experiment, you may be required to save the pellet, the supernatant, or both. ...
Showing: 861 - 880 of 916 results