NextSeq 500 paired end sequencing; Steroids-producing adrenocortical nodules (bulk RNA-seq)
Steroids-producing adrenocortical nodules (bulk RNA-seq)
To isolate nucleic acids, CPAs and periadrenal adipose tissue, as a control for germline DNA, were manually cut from the unstained FFPE sections. SPNs and adjacent adrenocortical samples as controls were microdissected from the unstained FFPE sections using a laser microdissection (LMD) microscope (Model LMD6500, Leica Microsystems) guided by the CYP17A1 immunohistochemistry slide. Genomic DNA and RNA were co-extracted from SPNs, CPAs, and adrenocortical tissue samples using the AllPrep DNA/RNA FFPE kit (Qiagen) according to the manufacturerâ€™s instructions. Libraries were prepared using the SMART-Seq Stranded Kit (Takara Bio, 634444).