We are working with patient-derived fresh tumor samples. We perform mechano-enzymatical digestion (DNAse + Colagenes II, 1h aprox., 37ºC) and send the single cell suspension to the scRNA-seq facility right after.

- Any evidence on the best digestion protocol?

- If the sample cannot be sent the same day, is it advisable to freeze the tumor disaggregate? should we perform an overnight digestion (at room temperature) instead?

- Which media do you use to run the sample?

Any advise will be of great help! Thanks