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WB-Validated RRM1 shRNA Lentiviral Particles#V9651

WB-Validated RRM1 shRNA Lentiviral Particles#V9651

$599.00Price

shRNA Knockdown Validated by WB

  • Aliases

    Ribonucleotide Reductase Catalytic Subunit M1; Ribonucleoside-Diphosphate Reductase Large Subunit; Ribonucleoside-Diphosphate Reductase Subunit M1; Ribonucleotide Reductase M1 Polypeptide; EC 1.17.4.1; RR1; Ribonucleotide Reductase Large Subunit; Ribonucleotide Reductase, R1 Subunit; RIR1; R1

  • Background

    NCBI Gene Entry: 6240

  • Tested Cell Line

    HeLa

  • Validation Methods

    RT-qPCR

    Western blotting (WB)

  • Shipping

    Shipped with dry ice. Immediately store the product in a standard freezer at -80°C upon receipt.

  • Storage

    Stored at -80°C for 1 year.

  • Instructions For Use

    The following protocol uses HeLa cell as an example assuming your cell culture medium is DMEM-based.

    1. Release 0.5 million HeLa cells into a 35 mm tissue culture dish in 2 mL of the growth medium (DMEM containing 10% FBS and 1% pen/strep). Cell density should reach 50-60% confluence the following day.
    2. 24 h after cell release, pre-warm the shRNA lentiviral medium to 37°C.
    3. Discard 1 mL of the original growth medium of the 35 mm dish.
    4. Using a serological pipette, gently mix the lentiviral solution 3 times.
    5. Carefully add 1 mL of the lentiviral solution to the well. Tip: To prevent splashing, add the solution to the dish along the wall.
    6. Add a polybrene stock solution to the culture medium at a final concentration of 5 μg/mL. Gently swirl the dish to mix.
    7. 48 h after cell release, without discarding the original medium, add another 1 mL of lentiviral medium directly into the dish.
    8. Add an additional polybrene stock solution into the dish to obtain a final concentration of 5 μg/mL. Tip: Now, the medium in the dish should be a total of 3 mL.
    9. 72 h after cell release, cells may reach confluence. Trypsinize the cells off the 35 mm dish and culture those cells in a 60 mm dish.
    10. Add puromycin to the dish at a final concentration of 4 μg/mL. Tip: To assess the efficacy of puromycin selection, culture a dish of wild-type HeLa cells as a negative control.
    11. Allow puromycin selection for 48 h. Almost all wild-type HeLa cells should die, while the dish infected with lentiviruses should have some remaining cells.
    12. Replace the medium with regular growth medium without puromycin and allow the cells to grow to confluence before harvesting or staining.
  • Note

    This product is for research use only. 

  • Data Sheet

    #V9651 RRM1 DS

Support

Orders

sales@genuinbiotech.com

Technical

info@genuinbiotech.com

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