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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 568 results for: mme

Showing: 541 - 560 of 568 results

  1. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...new window) NEB for more information). Thus, we recommend that you add 3-6 bases upstream of your restriction...important to digest plenty of starting material. We recommend using your entire PCR reaction and 1μg of recipient...space to cut out the bands. Because of this we recommend that you use a wide gel comb, run the gel on the...fuse your insert to your recipient plasmid. We recommend around 100ng of total DNA in a standard ligation...

  2. Plasmid Modification by Annealed Oligo Cloning (with Protocols)

    Type
    Protocol
    ...purify your DNA away from the agarose using a commercially available kit or standard protocol. Anneal oligos...EDTA) and mixed in equimolar concentrations. We recommend mixing 2μg each in a total volume of 50μL - add...oligos in a PCR tube. Place tube in a thermocycler programmed to start at 95°C for 2 minutes. Then, gradually...

  3. Protocol - Bacterial Transformation

    Type
    Protocol
    ...For the highest transformation efficiency, we recommend that you follow the instructions that came with...with your competent cells. Pro-Tips Commercial competent cells range significantly in their transformation...containing the appropriate antibiotic. Pro-Tips We recommend that you plate 50 μL on one plate and the rest...

  4. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...to use of your purified virus. Nonetheless, we recommend performing a buffer exchange before using the ...QuickSeal tube at the bottom using an 18 ga needle. Immediately start collecting 0.5 to 1 mL fractions in microcentrifuge...that has settled at the bottom of the column. We recommend concentrating to a minimum of 500 µL. If the concentrate...

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

    Type
    Protocol
    ...important to digest plenty of starting material. We recommend 1.5-2μg of donor plasmid and 1μg of recipient ...space to cut out the bands. Because of this we recommend that you use a wide gel comb, run the gel on the...fuse your insert to your recipient plasmid. We recommend around 100ng of total DNA in a standard ligation...

  6. AAV ddPCR Titration

    Type
    Protocol
    ... reduce the risk of contaminating reagents we recommend making small aliquots of master mixes, primers...Buffer containing 0.05% Poloxamer 188: Prepare immediately before use and vortex before using. 500 µL of... > 10 to be invalid. To reduce NTC values, we recommend wiping down all pipettes and equipment with 10%...

  7. Lentivirus ddPCR Titration

    Type
    Protocol
    ...of each sample. Samples can be used for ddPCR immediately or stored at -20 °C until ready to use. Preparing...infectious titer. Tips and Troubleshooting We recommend wiping down all pipettes and equipment with 10%... reduce the risk of contaminating reagents we recommend making small aliquots of master mixes, and primers...

  8. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ...given sequence. For a list of the commonly used commercially available restriction enzymes, see New England...aliquot of your plasmid of interest (see below for recommended amounts) Appropriate restriction enzyme (see ...

  9. Fluorescence Titering Assay

    Type
    Protocol
    ...Titer will vary between cell lines. It is not recommended to subject lentiviral supernatants to multiple...you are freezing and aliquoting virus, it is recommended that you titer from the frozen stock to account...

  10. Protocol - How to Inoculate a Bacterial Culture

    Type
    Protocol
    ...antibiotic powder can be dissolved in dH 2 0. Addgene recommends making 1000X stock solutions and storing aliquots...Antibiotic Concentrations Commonly Used Antibiotics Recommended Concentration Ampicillin 100 µg/mL Bleocin 5 ...

  11. Lab Safety for Biosafety Levels One and Two

    Type
    Protocol
    ... be done underneath a hood. Clean any spills immediately, and decontaminate as necessary. For large spills...bloodborne pathogens training. It is strongly recommended that anyone participating in BSL-2 work receives...

  12. Protocol - How to Ligate Plasmid DNA

    Type
    Protocol
    ...digestion . Most restriction enzymes digest DNA asymmetrically across their recognition sequence, which results... 1 vector molar ratio will work just fine. We recommend around 100ng of total DNA in a standard ligation...

  13. Immunocytochemistry

    Type
    Protocol
    ...between products. Addgene does not endorse or recommend specific products or equipment. Inclusion of this...antibody concentrations, incubation times, and recommended compatible reagents. Secondary antibodies must...

  14. Colony Formation Titering Assay

    Type
    Protocol
    ...Titer will vary between cell lines. It is not recommended that lentiviral supernatants be subjected to ...you are freezing and aliquoting virus, it is recommended that you titer from the frozen stock to account...

  15. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...between products. Addgene does not endorse or recommend specific products or equipment. Inclusion of this...water IgG isotype standard 2.5 mg/mL - can use commercial standard or validate an in-house standard Before...

  16. Western Blot

    Type
    Protocol
    ...between products. Addgene does not endorse or recommend specific products or equipment. Inclusion of this...a clean microcentrifuge tube. Use the lysate immediately or store at -80 °C until ready to use. Section...

  17. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...between products. Addgene does not endorse or recommend specific products or equipment. Inclusion of this...wash, aspirate the wash buffer from the wells. Immediately before use, mix 5 mL TMB Solution and 5 mL Peroxide...

  19. Protocol - How to Streak a Plate

    Type
    Protocol
    ...setting is important for organization, and it is recommended that you keep a standard labeling system for ...
Showing: 541 - 560 of 568 results