This protocol is used to generate and quantify cDNA from miRNA and mRNA in frozen tissue samples. Samples are prepared by immediate submersion in RNALater and incubated at room temp overnight, drained, snap frozen, and stored at -80ºC. RNA is extracted from these tissues using the Qiagen miRNeasy kit. This kit extracts total RNA, despite it's name. This total RNA is used as input for the miScript II RT Kit, which will convert the RNA to cDNA. The kit has two buffers, we use the HiFlex Buffer, which is designed to capture both miRNA and mRNA. The cDNA generated from the miScript II RT Kit is then split into two aliquots. One aliquot is used for detecting and quantifying mature miRNA, while the second aliquot is used for detecting and quantifying mRNA. To quantify mature miRNA, we use the miScript Primer Assay Kit, followed by the miScript SYBR Green PCR kit. For detecting an quantifying mRNA, we use the QuantiTect Primer Assay, followed by the QuantiTect SYBR Green PCR Master Mix (provided in the miScript SYBR Green PCR kit).