Quantitative Analysis of SULT2B1 Expression

Total RNA was isolated using TRIzol Reagent (Thermo Fisher) according to the manufacturer's protocol. cDNA was synthesized with M-MuLV Reverse Transcriptase (New England Biolabs, Ipswich, MA) as described previously.^{18 (link)} Quantitative real-time polymerase chain reaction (qRT-PCR) was conducted on LightCycler 96 (Roche, Basel, Switzerland) using PerfeCTa qPCR FastMix II (Quantabio, Beverly, MA) according to the manufacturer's protocol. PrimeTime qPCR Assay probes specific for human SULT2B1b (Hs.PT.58.22508626) or for mouse Sult2b1 (Mm.PT.58.5982373) (Integrated DNA Technologies, Skokie, IL) were used and the transcript levels were normalized using TaqMan Ribosomal RNA Control Reagents (Thermo Fisher).

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Yang J., Broman M.M., Cooper P.O., Lanman N.A., Strand D.W., Morrissey C, & Ratliff T.L. (2019). Distinct expression patterns of SULT2B1b in human prostate epithelium. The Prostate, 79(11), 1256-1266.

Publication 2019

TRIzol Reagent M-MuLV Reverse Transcriptase LightCycler 96 PerfeCTa qPCR FastMix II TaqMan Ribosomal RNA Control Reagents

Assay Cdna Human M mulv Mouse Quantitative real time polymerase chain reaction Reverse transcriptase Ribosomal rna Trizol

Corresponding Organization : University of Washington

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Transcript levels of human SULT2B1b
Transcript levels of mouse Sult2b1

control variables

Total RNA isolation using TRIzol Reagent
CDNA synthesis with M-MuLV Reverse Transcriptase
Quantitative real-time PCR using LightCycler 96 and PerfeCTa qPCR FastMix II
Normalization using TaqMan Ribosomal RNA Control Reagents

controls

Positive control: None mentioned
Negative control: None mentioned

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