Protocol detail

Quantification of SDHB Expression and Editing

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SDHB gene expression and c.136C>U RNA editing was quantified by RT–PCR16 (link). Unless noted otherwise, total RNA, genomic DNA and plasmid DNA were isolated using material and methods provided with TRIzol, DNA Wizard Genomic DNA Purification Kit (Promega) and Plasmid Kit (Qiagen, Germantown, MD), respectively. RNA/DNA was quantified by spectrophotometry on a Nanodrop 2000 instrument (Thermo Fisher). Proteins were quantified using Bio-Rad Dc assay with BSA standards. Statistical tests were two-tailed and were performed using R 3.0, Excel 2010 (Microsoft, Redmond, WA), or Prism 6.0 (GraphPad, San Diego, CA) software.

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Sharma S., Patnaik S.K., Thomas Taggart R., Kannisto E.D., Enriquez S.M., Gollnick P, & Baysal B.E. (2015). APOBEC3A cytidine deaminase induces RNA editing in monocytes and macrophages. Nature Communications, 6, 6881.

Publication 2015

DNA Wizard Genomic DNA Purification Kit Plasmid Kit Nanodrop 2000 instrument Dc assay Prism 6.0

Assay Gene expression Genomic Plasmid Prism Promega Proteins Sdhb Spectrophotometry Trizol

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Variable analysis

independent variables

SDHB gene expression

dependent variables

C.136C>U RNA editing

control variables

Total RNA, genomic DNA and plasmid DNA were isolated using material and methods provided with TRIzol, DNA Wizard Genomic DNA Purification Kit (Promega) and Plasmid Kit (Qiagen, Germantown, MD), respectively
RNA/DNA was quantified by spectrophotometry on a Nanodrop 2000 instrument (Thermo Fisher)
Proteins were quantified using Bio-Rad Dc assay with BSA standards

controls

Positive controls not specified
Negative controls not specified

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