Optimized protocol for the RNA isolation of laser microdissected formalin-fixed paraffin-embedded uterine scar tissues for RNA expression analyses v1

Samples for histological analyses are often formalin-fixed paraffin-embedded (FFPE) and slide-mounted. This complicates RNA extraction for molecular downstream applications. Additionally, when the region of interest is limited to a smaller area with laser microdissection (LMD), extracting adequate quality and quantity RNA is difficult. This is a protocol for maximized RNA output from FFPE tissue devised to identify and analyze gene expression of human maternal uterine scar tissue obtained from uterotomy scars resulting from prior cesarean deliveries. Gomori Trichrome staining allowed for region identification for LMD. Successful RNA isolation, reverse transcription, and quantitative real-time polymerase chain reaction (qRT-PCR) were performed. This description provides an optimized step-by-step protocol that yields sufficient RNA for qRT-PCR analyses from challenging tissue such as LMD-FFPE samples.