Protocol: Harvesting RNA from Cells Cultured on P3D Scaffolds

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Application

This protocol explains how to harvest RNA from cells cultured on P3D Scaffolds. For more protocols, please visit Ossiform P3D Scaffolds product page.

Materials

  • Qiazol (or other RNA harvesting agent)
  • Shaking table
  • RNA-clean Eppendorf tubes
  • Vortexer
  • Ice

Flowchart

Flowchart of protocol for harvesting RNA from cells cultured on Ossiform P3D Scaffolds

Notes before starting and general advice on material handling

  • All handling of the P3D Scaffolds products should be performed using gloves, according to the standard aseptic methods.
  • The scaffolds are supplied sterile by dry heat sterilization and remains sterile until opened.

Procedure

  1. Aspirate cell medium.
  2. Add 450-500 uL Qiazol to each well.
  3. Incubate 2-10 min at RT on shaking table at 900 rpms. Increasing time can increase yield.
  4. Flush the scaffold surface by pipetting and transfer lysate to RNA-clean Eppendorf tubes on ice.
  5. Mix by pipetting the full volume up and down.
  6. Vortex each sample for 10 sec and incubate at RT for 5 min.
  7. Store samples at -80ºC or continue with downstream procedures.

Disclaimer

The products are “For Research Use Only (RUO)” and should not be used for clinical, diagnostic, or therapeutic purposes. Ossiform and its distributor, Ilex Life Sciences LLC, make no other warranties, expressed or implied, including the implied warranty of merchantability and the implied warranty of fitness for particular purpose.

Acknowledgement

This protocol has been adapted from the protocol published by Ossiform. Ilex Life Sciences is an authorized distributor of Ossiform P3D Scaffolds research product line.

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