Total RNA was extracted from human cells using RNAiso Plus reagent (9109, Takara). cDNA was synthesized using a reverse transcriptase kit (RT200, Enzynomics). RT-qPCR was performed with SYBR Green (RT500M, Enzynomics) using a Real-Time System (Bio-Rad, CFX96TM). All data were normalized to glyceraldehyde-3-phosphate dehydro-genase expression. Information on the primer sets (Biosearch Technologies, Novato, CA) used in the present study is listed in Table 2 .
Rt200
Manufactured by Enzynomics
The RT200 is a real-time PCR (Reverse Transcription Polymerase Chain Reaction) instrument designed for nucleic acid amplification and detection. The core function of the RT200 is to perform sensitive and precise quantitative analysis of DNA and RNA samples.
Automatically generated - may contain errors
2 protocols using rt200
1
Total RNA Extraction and RT-qPCR Analysis
2
Quantitative Gene Expression Analysis
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Total RNA was extracted from human cells using TRIzol reagent (Ambion) as described by the manufacturer. cDNA was then synthesized using reverse transcriptase kits (RT200, Enzynomics). Fragments were amplified by qRT‐PCR using primers (Table 1 ), SYBR Green (RT500M, Enzynomics), and the CFX96™Real‐Time System (Bio‐Rad Laboratories Inc.). The relative mRNA expression of target genes was calculated using the comparative CT method. All target genes were normalized to GAPDH in multiplexed triplicate reactions. Differences in CT values were calculated for each target mRNA by subtracting the mean value of GAPDH (relative expression = 2−ΔCT).
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