Total RNA was extracted from control and treated cells using TRIzol reagent (Life Technologies, Inc., Carlsbad, CA) according to manufacturer instructions and the cDNA synthesis was carried out as described by us earlier (29 (link)). About 2 µg of template cDNA was used for cycling in a thermal cycler (Thermo Fisher, Pittsburgh, PA). The PCR products were separated on a 1.5% agarose gel, stained with 0.5 mg/mL ethidium bromide, and visualized under UV light using BioRad Versa Doc imager.
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RNA Extraction and cDNA Synthesis
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Corresponding Organization : Texas Tech University
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Variable analysis
independent variables
- Control cells
- Treated cells
- Total RNA expression
- CDNA synthesis
- PCR product separation and visualization
- TRIzol reagent (Life Technologies, Inc., Carlsbad, CA) used for total RNA extraction
- CDNA synthesis carried out as described in previous work (29)
- About 2 µg of template cDNA used for cycling in a thermal cycler (Thermo Fisher, Pittsburgh, PA)
- PCR products separated on a 1.5% agarose gel, stained with 0.5 mg/mL ethidium bromide, and visualized under UV light using BioRad Versa Doc imager
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