Quantitative RNA Expression Analysis

RNA extraction and real-time quantitative PCR (qPCR) were carried out using procedures that we had previously described (12 (link)). Briefly, total RNA was extracted using a PureLink RNA minikit (Life Technologies). DNase (Ambion)-treated RNA samples were analyzed on a denaturing formaldehyde agarose gel for assessing purity, quality, and concentration. A SuperScript III cDNA synthesis kit (Life Technology) was used for the first-strand cDNA synthesis following the manufacturer’s protocols. Constitutively expressed housekeeping gene TEF1 was used as an endogenous control for the normalization of expression of the other genes studied.

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Zhao Y., Upadhyay S, & Lin X. (2018). PAS Domain Protein Pas3 Interacts with the Chromatin Modifier Bre1 in Regulating Cryptococcal Morphogenesis. mBio, 9(6), e02135-18.

Publication 2018

PureLink RNA minikit DNase SuperScript III cDNA synthesis kit

Agarose Cdna Dnase Formaldehyde Genes Housekeeping gene Real time pcr Synthesis

Corresponding Organization :

Other organizations : Texas A&M University, University of Georgia

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Expression of the genes studied

control variables

Constitutively expressed housekeeping gene TEF1 used as an endogenous control for normalization of gene expression

controls

Positive control: None mentioned
Negative control: None mentioned

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