RNA immunoprecipitation–quantitative polymerase chain reaction

TRIzol (Invitrogen, Carlsbad, CA) was used to extract total RNAs from the immunoprecipitate of Csde1 according to the manufacturer’s instructions. Random primers were then used for complementary DNA (cDNA) synthesis. To detect whether Csde1 target genes were significantly and specifically enriched in the Csde1 immunoprecipitate, we used IgG RNA as a reference and performed quantitative reverse transcription polymerase chain reaction (RT-PCR) to determine the relative level of specific RNAs in the IgG and Csde1 immunoprecipitates. Quantitative PCR (qPCR) data represent the mean values from at least three independent experiments. Various genes were selected for PCR amplification in both Csde1 and IgG immunoprecipitates.

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