GSM9025875: 2393-XX-90; Homo sapiens; RNA-Seq
Samples were incubated in an optimized lysis buffer, which contains proteinase K, to release RNA from the sample sections. This was followed by an incubation at a higher temperature to partially reverse formalin crosslinking of the released nucleic acids and improve RNA yield and quality. This was followed by DNase treatment that was optimized to eliminate all genomic DNA, including very small DNA fragments. The lysate was then mixed with Buffer RBC, and ethanol was added to provide appropriate binding conditions for RNA. The sample was then applied to an RNeasy MinElute spin column, where the total RNA bound to the membrane and contaminants were effectively washed away. RNA was then eluted in 30µl of RNase-free water. Libraries were generated using NovaSeq.