Fig. 7
From: Advancing brain barriers RNA sequencing: guidelines from experimental design to publication
Overview of commonly used RNA isolation protocols. Preparation of BBB-derived samples according to the type of sample. Cells in suspension are first collected by centrifugation, while adherent cultured cells are commonly trypsinized; then lysis buffer is added and cells are homogenized before proceeding to isolation of the RNA. Fresh frozen tissue can be mechanically disrupted in lysis buffer; debris should be removed by centrifugation before RNA isolation. Formalin-fixed paraffin-embedded (FFPE) tissue is first deparaffinized, and tissue disruption can be achieved by enzymatic (proteinase K) and/or mechanical means; de-crosslinking is followed by addition of lysis buffer, and then RNA is isolated