Some important points includePlease find below a list of key points to consider to maximize your RNA extraction:
Remove as much paraffin as possible around the sample before beginningtissue.
If you have a tissue with a small surface area, you can consider reducing the amount of the FFPE Lysis Buffer (minimum of 70μldown to 70 μl). Please see this FAQ How can I increase my final yield? for more information.
Check that the sample is well submerged in the FFPE Lysis Buffer.
The FFPE Removal Solution solution is light sensitive and provided in a brown bottle. Take care when storing/using this reagent.Ensure Spin Column is Please carefully follow the storage and handling recommendations described in the User Guide.
Ensure the FFPE SPLIT RNA columns are prepared appropriately. Please vortex well and spin down prior to removing the Storage Buffer, and ensure no air bubbles are present. Refer to this FAQ Please see the following FAQ: Do you have any specific recommendations for spin column preparation? for additional tips on column preparation.
Prior to DNase digestion, check for the absence presence of paraffin in the aqueous upper phase when letting the sample cooldown. If you observe a jelly gelatinous layer on top of the aqueous phase, this means that you have some paraffin left in your sample. Reheat In this case, please re-incubate the sample at 60°C and centrifuge again. Take out the aqueous phase.
Increase the centrifugation speed to maximize the phase separation.
When centrifuging the final eluate, do not increase the speed. If When >1,000 × g is used, this can lead to impurities impurities could be present in your final sample.