Total RNA from cells was extracted using TRIzol (Invitrogen). Two microgram of RNA was reverse-transcribed using M-MLV reverse transcriptase (Promega) and cDNA was resuspended in 40 μl of RNase-free water. PCR was then performed with 2μl of cDNA, using the conditions and primers described in Supplemental Material .
Protocol detail
Extraction and Reverse Transcription of Total RNA
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Corresponding Organization :
Other organizations : Boston University, National Institutes of Health, Universität Ulm, National Heart Lung and Blood Institute
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Variable analysis
independent variables
- None explicitly mentioned
- Gene expression changes
- RNA extraction method (TRIzol)
- Reverse transcription method (M-MLV reverse transcriptase)
- PCR conditions and primers (as described in Supplemental Material)
- Positive control: Not specified
- Negative control: Not specified
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