Protocol tips
Upstream tips |
Protocol tips |
Downstream tips |
The Qubit® assays are designed to be performed at room temperature, as temperature fluctuations can influence the accuracy of the assay. |
Dilute Qubit® RNA HS Reagent 1:200 in Qubit® RNA HS Buffer. |
|
Upstream tips |
The Qubit® assays are designed to be performed at room temperature, as temperature fluctuations can influence the accuracy of the assay. |
Protocol tips |
Dilute Qubit® RNA HS Reagent 1:200 in Qubit® RNA HS Buffer. |
Publication protocol
Quantitative reverse transcription-PCR was performed using total RNA with specific primers and probes (S3 Table) designed using Primer Express software v3.0.1 (Life Technologies) essentially as previously described [16] except that a BioRad CFX Real-Time System was also used for analyses. TaqMan probes (Biosearch Technologies) were used for gene quantification using TaqMan® RNA-to-CT 1-Step Kit (Life Technologies). Gene expression data were normalized to level of cyclophilin b (Ppib), or TATA box binding protein (Tbp). In addition, the total nanograms of input RNA/reaction was quantified using Qubit HS RNA Assay Kit (Life Technologies) and can be used to normalize gene expression by RNA input if necessary.
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Diet-induced adipose tissue expansion is mitigated in mice with a targeted inactivation of mesoderm specific transcript ()
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