2.2. Extraction and Reverse Transcription of Mouse Gastric Tissue RNA and PCR Amplification of MIC-1 Gene Sequence

Healthy mouse gastric tissue was isolated, and total RNA was extracted using Trizol (9108). RNA integrity was assessed using a 1% agarose gel. The intact RNA was reverse transcribed into cDNA using 5x Primescript RT Master Mix. The MIC-1 gene sequence was obtained from the GenBank database, and primers for MIC-1 were designed with QuickCut HindIII (1,615) and QuickCut EcoRI (1,605) restriction endonuclease sites in the forward and reverse primers, respectively. The PCR reaction mixture contained GoTaq qPCR Master Mix (M722), forward and reverse primers, MIC-1 DNA, and sterile water. The resulting PCR products were analyzed by agarose gel electrophoresis.