RNeasy Plant Mini Kit

RNA isolation / purification Yeast - Neurospora crassa

Experiment
RNA isolation / purification Yeast - Neurospora crassa
Product
RNeasy Plant Mini Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

Protocol tips
Both enzymatic and mechanical lysis can be used for yeast. Enzymatic lysis doesn't require additional lab equipment, but takes longer than mechanical disruption.

Publication protocol

Cultures were grown at 30° with constant shaking for 12 hr (in the absence of Cm) or 14 hr (in the presence of Cm). RNA was isolated using the QIAGEN (Ontario, Canada) RNeasy Plant Mini Kit. RNA integrity was measured using an Agilent 2100 Bioanalyzer system and the Agilent RNA 6000 Nano kit (Agilent Technologies). Only those samples that had an RNA integrity number value of >9 were used for cDNA synthesis. First-strand cDNA was synthesized using SuperScript III Reverse Transcriptase (Invitrogen). Reactions were performed using KAPA SYBR Fast qPCR reagents (KAPA Biosystems) and the StepOnePlus qPCR system (Applied Biosystems, Foster City, CA). Data were analyzed using StepOne software (Applied Biosystems). The β-tubulin gene was the internal control for ΔΔCt quantification. The qPCR data for each gene examined were obtained from four biological replicates, each with three technical replicates.

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Papers

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Paper title
Alternative Oxidase Transcription Factors AOD2 and AOD5 of Control the Expression of Genes Involved in Energy Production and Metabolism
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Manufacturer protocol

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