Total RNA (500 ng) was DNAase treated, and complementary DNA was generated using the iScript complementary DNA synthesis kit (Bio-Rad Laboratories, Hercules, CA). qPCR was performed with the Applied Biosystems Incorporated 7900HT Fast Real-Time System (Life Technologies, Grand Island, NY) and LightCycler FastStart DNA Master SYBR Green as a ready-to-use reaction mix with ROX (Roche, Basel, Switzerland). Relative expression was calculated using a standard curve method, as described.39 (link),40 Results were normalized to GAPDH and fold induction compared with controls. cDNAs were amplified using TaqMan Gene Expression Assay (Hs00226845_m1) and primers (Integrated DNA Technologies, Coralville, IA) (Supplementary Table 1 ).41 (link)
Taqman gene expression assay hs00226845 m1
Manufactured by Integrated DNA Technologies
The TaqMan Gene Expression Assay (Hs00226845_m1) is a pre-designed real-time PCR assay developed by Integrated DNA Technologies. It is used for the quantitative analysis of gene expression levels.
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2 protocols using taqman gene expression assay hs00226845 m1
1
Quantitative Real-Time PCR Assay Protocol
2
Quantitative Real-Time PCR Assay Protocol
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Total RNA (500 ng) was DNAase treated, and complementary DNA was generated using the iScript complementary DNA synthesis kit (Bio-Rad Laboratories, Hercules, CA). qPCR was performed with the Applied Biosystems Incorporated 7900HT Fast Real-Time System (Life Technologies, Grand Island, NY) and LightCycler FastStart DNA Master SYBR Green as a ready-to-use reaction mix with ROX (Roche, Basel, Switzerland). Relative expression was calculated using a standard curve method, as described.39 (link),40 Results were normalized to GAPDH and fold induction compared with controls. cDNAs were amplified using TaqMan Gene Expression Assay (Hs00226845_m1) and primers (Integrated DNA Technologies, Coralville, IA) (Supplementary Table 1 ).41 (link)
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