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Addgene

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

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We narrowed to 21 results for: SUB

Showing: 1 - 20 of 21 results

  1. Plasmid Cloning by Restriction Enzyme Digest (with Protocols)

    Type
    Protocol
    ...enzyme digest (subcloning), including design and experimental procedures.... Protocols Subcloning Plasmid Cloning...Cloning by Restriction Enzyme Digest (aka Subcloning) You may also like... Restriction Digest of Plasmid...out our Empty Backbone Reference . Background Subcloning by restriction digest is a commonly used lab ...YGOI is in a bacterial expression vector. Using subcloning, you can easily move YGOI into a mammalian expression...find two different restriction enzymes for your subcloning. It is also possible to use a single enzyme, ...

  2. DNA Quantification

    Type
    Protocol
    ... of a substance in that liquid. In order to accurately measure the concentration of a substance based ...to determine the concentration of a particular substance in that liquid. Molecules absorb different wavelengths...need to know the wavelength of light that your substance maximally absorbs. In the case of nucleic acid...

  3. Western Blot

    Type
    Protocol
    ...powder 96-well microtiter plate Chemiluminescence substrate Plastic wrap Primary antibody Secondary antibody...absorbance of the duplicate samples on the plate. Subtract the average absorbance of the blank from all of...shaking platform. Prepare the chemiluminescence substrate by mixing 1:1 reagent A to reagent B. Gently incubate...

  4. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...,5'-tetramethyl-[1,1'-biphenyl]-4,4'-diamine) substrate kit, Pierce 34021 Tween-20, VWR 0777-1L Foil Before...Solution and 5 mL Peroxide Solution from the TMB substrate kit. Cap the tube and invert several times to ...secondary antibody for 2 hours, reacted with TMB substrate and the reaction stopped after 15 minutes. Absorbance...

  5. General Transfection

    Type
    Protocol
    ...function. We specifically use this protocol with a subclone of HEK293T cells optimized for viral production...Workflow Timeline Day 0: Seed HEK293T cells (or a subclone of HEK293T optimized for viral production) Day...

  6. Pipetting Protocol

    Type
    Protocol
    ... lower the pipette tip into the liquid. Do not submerge the pipette itself into the liquid and avoid touching...contaminating the contents. Once the end of the tip is submerged, slowly release the plunger, allowing it to go...

  7. Pouring LB Agar Plates

    Type
    Protocol
    ... a water bath at 60 ℃ with sufficient water to submerge ~75% of the bottle containing your molten gel ...removing anything from the autoclave. Partially submerge your molten gel-mix in the 60 ℃ water bath. Pro-Tip...

  9. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...can be converted into lentiviral particles for subsequent infection of a target cell line. Once introduced...toxic to some cell lines. In these cell lines, substitute protamine sulfate for polybrene. c. Add lentiviral...

  12. Weighing Reagents Protocol

    Type
    Protocol
    ...weighing and prevent cross-contamination with other substances. They also help you transfer the material to ...

  13. Ligation Independent Cloning

    Type
    Protocol
    ...buffer) to ensure optimal salt concentrations in subsequent reactions. Step 3: Create Vector Overhangs Treat...

  14. Fluorescence Titering Assay

    Type
    Protocol
    ...vary between cell lines. It is not recommended to subject lentiviral supernatants to multiple freeze-thaw...

  15. Gibson Assembly Protocol

    Type
    Protocol
    ... DNA 5’ end. Complementary DNA fragments can subsequently anneal to each other. Phusion High-Fidelity ...

  16. Colony Formation Titering Assay

    Type
    Protocol
    ...not recommended that lentiviral supernatants be subjected to multiple freeze-thaw cycles. This protocol ...

  20. CRISPR Library Amplification

    Type
    Protocol
    ...incremental, ordered cell lysis. E. coli cells are subject to lysis by freeze-thaw if not suspended in cryoprotectant...
Showing: 1 - 20 of 21 results