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Do you have any specific recommendations for FFPE SPLIT RNA columns preparation?

Please find below a list of recommendations to optimize your column preparation:

  • Vortex the columns upside-down to homogenize the matrix. Place the columns in a 2 ml collection tube in the upright position at least 1 hour before starting the protocol to ensure the matrix sediments well. Alternatively, columns can also be prepared (homogenization and upright position) upon receiving the kit or the day before starting the experiment.

  • Before beginning, check that there are no air bubbles in the matrix. If so, flick or gently spin down the columns until it is free from air bubbles. Presence of air bubbles may lead to impurities in the RNA sample.

  • Open the lid of the column and centrifuge at 1,000 × g.  Do not exceed 1,000 × g! Higher centrifugation speeds can dry out the column, which will result in suboptimal elution and lower yields.

  • Ensure the matrix is not highly titled in the column. If using a fixed rotor for centrifugation, a titled angle ~30° is expected to be observed. Higher angles may alter the purification ability leading to presence of impurities and/or lower yields. If the angle is too high, we recommend to resuspend the matrix by vortexing and leave the column overnight to sediment before conditioning. If interested, Column Adapters for Plate Centrifuges with swing rotors are available to ensure matrix is horizontal and not tilted (Cat. No. 239).

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