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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 14 results for: PIR

Showing: 1 - 14 of 14 results

  1. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...primary antibody to use will vary and needs to be empirically determined. If possible, run an initial test ...plate seal from the 96-well plate and set aside. Aspirate the wells and use a multichannel pipette to add...a total of three washes. After the final wash, aspirate the wash buffer from the wells. Prepare the blocking...a total of three washes. After the final wash, aspirate the wash buffer from the wells. Prepare antibody...antibody concentration will vary and needs to be empirically determined. We suggest starting with three concentrations...

  2. Lentivirus ddPCR Titration

    Type
    Protocol
    ... Harvest Gently aspirate media from the 6-well plates. Be sure to use a new aspirating pipette for each... 37 °C for 30 min. Gently aspirate media from wells, using a new aspirating pipette for each well. Wash...Wash wells with 1 mL of 1X PBS and aspirate, using a new aspirating pipette for each well. Detach cells...Equipment Class II, Type A2 Biological Safety Cabinet Aspirating unit Microcentrifuge, Eppendorf, 5425R Droplet...tube. Centrifuge for 5 min at 100 x g . Gently aspirate supernatant. Wash cell pellets in 500 µL PBS. .... Centrifuge for 5 min at 100 x g . Gently aspirate supernatant. Extract genomic DNA according to the ...

  3. Colony Formation Titering Assay

    Type
    Protocol
    ... wells, aspirate the media from the wells. Gently wash the cells with 1 mL of PBS and aspirate wash. Filter...required to kill your target cell line needs to be empirically determined. Treat the target cells with a range...added until 48–72 h after transduction. Gently aspirate the media from the cells. Add 1.5 mL of DMEM complete...colonies should be visible. Every 3–4 days, gently aspirate the media and replace it with fresh DMEM complete...

  4. General Transfection

    Type
    Protocol
    ...optimal mass DNA:mass PEI ratio will need to be empirically determined for each new batch of 1 mg/mL PEI ... the cells at 37 °C, 5% CO 2 for ~20 h. Gently aspirate media, add 10 mL fresh DMEM complete containing...Pro-Tip The ratio of µg DNA:µg PEI needs to be empirically determined. Once a batch of PEI is prepared, ...following morning. The following morning, carefully aspirate the media. Replace the media with 15 mL of DMEM...

  5. Lentivirus Production

    Type
    Protocol
    ...optimal mass DNA:mass PEI ratio will need to be empirically determined for each new batch of 1 mg/mL PEI ...therefore the ratio of μg DNA:μg PEI needs to be empirically determined. Once a batch of PEI is prepared, ...the media and chloroquine and mix well. Gently aspirate the media out of the previously seeded 10 cm plate...following morning. The following morning, carefully aspirate the media. Replace the media with 10 mL of DMEM...

  6. AAV Production in HEK293 Cells

    Type
    Protocol
    ...each T-175 flask: Aspirate culture media and rinse once with 10 mL of PBS. Aspirate PBS and add 2 mL of...at ~80% confluence. Aspirate culture media and rinse once with 60 mL PBS. Aspirate PBS and add 35 mL of...

  7. Fluorescence Titering Assay

    Type
    Protocol
    ...Untransduced 0 1498.5 1.5 Mix the dilutions well Gently aspirate media from the cells. Add 1.5 mL of a viral dilution...calculating the titer. Incubate for 48–72 h. Gently aspirate media and replace with 1 mL of PBS. Calculate ...

  8. AAV ddPCR Titration

    Type
    Protocol
    ...generated, use a 20–200 µL multichannel pipet to aspirate 40 µL of droplets. Pro-Tip To ensure that the ...45° angle. Count to 20 while slowly and gently aspirating the droplets. Transfer the droplets to a prechilled...

  10. Immunocytochemistry

    Type
    Protocol
    ...Section 2: Fixing and permeabilizing cells Gently aspirate the media from the 24-well plate. Wash each well...
Showing: 1 - 14 of 14 results