1% for the planet logo
< Back to Support Article Search Results

Does CST have a protocol for treating cells with poly(dA:dT)?

The suggested protocol for treating cells with poly(dA:dT) can be found below: 

Poly(dA:dT) Treatment Protocol (5 μg/mL final concentration)

Reagents:

Procedure:

24 hours prior to transfection:
  • Plate cells in two 15 cm plates with 21 mL media with serum (no antibiotic)

On transfection day:

  1. Prepare two falcon tubes per plate to be transfected:
    • For mock: One tube labeled “FuGENE” and one tube labeled “Mock”
    • For poly(dA:dT): One tube labeled “FuGENE” and one tube labeled “Poly(dA:dT)”
  2. Add 4.5 mL of 1X serum-free medium to each tube.
  3. Add 90 μL of FuGENE 6 reagent to each “FuGENE” tube.
  4. Add 150 μL of poly(dA:dT) to the “Poly(dA:dT)” tube and add 150 μL of 1X PBS to the “Mock” tube.
  5. Incubate for 20 min at room temperature.
  6. Combine the diluted reagent by adding the full volume from the “FuGENE” tube to the appropriate "Poly(dA:dT)" or "Mock" tube and mix gently.
  7. Incubate for 20 min at room temperature.
  8. Add FuGENE:DNA complex mixture dropwise to appropriately labeled plates.
  9. Incubate for the desired number of hours.
  10. Wash cells with 1X PBS, lyse with 1 mL 1X SDS + DTT lysate.

*Alternatively, untreated cells can be used as a negative control if a mock transfection is not necessary.

Last updated: April 25, 2024

Was this article helpful?

Technical Support

Email: [email protected]

Send Us a Message

Call: 877-678-8324

Customer Support

Email: [email protected]

Send Us a Message

Call: 877-616-2355

Fax: 866-432-6112

Contact Sales

Email: [email protected]

Send Us a Message

Call: 877-616-2355