Total RNA was isolated with the QIAamp RNA Isolation Kit (Qiagen, Hilden, Germany) and converted to cDNA using the AffinityScript QPCR cDNA Synthesis Kit (Agilent Technologies, Santa Clara, California). The cDNA was then used as the template for quantitative real-time polymerase chain reaction (qPCR) using Brilliant III Ultra-Fast QPCR Master Mix on Mx3005P QPCR system (Agilent technologies) and the following PrimeTime Predesigned qPCR Assays (Integrated DNA Technologies, Coralville, Iowa): human LDLR (Hs.PT.58.14599757), human GAPDH (Hs.PT.39a.22214836), human HMGCR (Hs.PT.58.41105492), human PCSK9 (Hs.PT.58.203171419), SREBP-2 (Hs.PT.56a.2651954) and human TFRC (Hs.PT.39a.22214826). qPCRs were run in duplicate and GAPDH was used as the normalizing gene. The 2−ΔΔCt method was used to calculate relative mRNA levels.
Brilliant 3 ultra fast qpcr master mix on mx3005p qpcr system
Manufactured by Agilent Technologies
Sourced in United States
The Brilliant III Ultra-Fast QPCR Master Mix is a reagent designed for use with the Mx3005P QPCR system. It is formulated to enable rapid and sensitive quantitative PCR (qPCR) analysis.
Automatically generated - may contain errors
2 protocols using brilliant 3 ultra fast qpcr master mix on mx3005p qpcr system
1
Quantitative PCR Analysis of Cholesterol Pathways
2
Quantitative gene expression analysis
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The total RNA was isolated with the QIAamp RNA Isolation Kit (Qiagen, Hilden, Germany) and converted to cDNA using the AffinityScript QPCR cDNA Synthesis Kit (Agilent Technologies, Santa Clara, CA, USA). The cDNA was then used as the template for quantitative real-time polymerase chain reaction (qPCR) using Brilliant III Ultra-Fast QPCR Master Mix on Mx3005P QPCR system (Agilent technologies, Santa Clara, CA, USA) and the following PrimeTime Predesigned qPCR Assays (Integrated DNA Technologies, Coralville, IA, USA): human LDLR (Hs.PT.58.14599757), human GAPDH (Hs.PT.39a.22214836), human ZFP36L1 (Hs.PT.56a.20479742), human ELAVL1 (Hs.PT.58.22593583), and human RPS6KA1 (RSK1) (hsPT58.1350004). qPCRs were run in duplicate and GAPDH was used as the normalizing gene. The 2−ΔΔCt method was used to calculate relative mRNA levels.
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