Abi prism 7900 high throughput real time pcr system

Total RNA was isolated from cells and reversely transcribed into cDNAs according to the supplier's protocols. We performed real-time PCR in an ABI Prism® 7900 High-Throughput Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) with gene specific primers (Table 1). The reaction condition was the same as our previous report [25 (link)]. For relative quantification of target genes, the comparative threshold cycle (C_T) method was used with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an internal control. The mRNA expression levels of target genes were plotted as fold of control cells, which were designated as 1.

After treatment, total RNA was isolated from BGC-823 or NCI-H87 gastric cancer cells and tumor tissue using RNA isolater Total RNA Extraction Reagent (Vazyme Biotech Co. Ltd) and reverse transcribed into cDNA using HiScript II Q Select RT SuperMix for qPCR (Vazyme Biotech Co. Ltd) according to the supplier’s protocol. Real-time PCR was performed with gene-specific primers (Table 1, Thermo Fisher Scientific Inc.) with 18s as internal control. The reactions were performed in an ABI Prism® 7900 High-Throughput Real-Time PCR System (Applied Biosystems, Foster City, CA, U.S.A.). The comparative threshold cycle (C_T) method was used for relative quantification of target gene expression, which was plotted as fold of control.