Qubit h 2.0 fluorometer

Samples (~100 mg) were ground in liquid nitrogen and then mixed with Trizol reagent (1 ml) and total RNA was extracted following the manufacture’s instructions (Invitrogen, Life Sciences). Genomic DNA in samples was digested using TURBODNase (Invitrogen) according to the manufacture’s instruction. Total RNA quality and quantity were analyzed by agarose gel electrophoresis and via NanoDrop 2000 spectrophotometric analyses. Quantification of RNA concentrations in samples was performed using a Qubit H 2.0 fluorometer (Invitrogen, Carlsbad, CA). Finally, 2 μg total RNA from different tissues were used to construct cDNA libraries using High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA) in terms of standard protocol.

Samples (∼100 mg) were ground in liquid nitrogen using a mortar and pestle, after which Trizol reagent (1 ml) was added, and total RNA extracted following the manufacturers’ protocols (Invitrogen, Carlsbad, CA, United States). Genomic DNA in samples were digested using TURBODNase (Invitrogen, Carlsbad, CA, United States). Total RNA quality and quantity were analyzed by agarose gel electrophoresis and via NanoDrop 2000 spectrophotometric analyses. Quantification of RNA concentrations in samples was performed using a Qubit H 2.0 fluorometer (Invitrogen, Carlsbad, CA, United States) and cDNA libraries were constructed using 2 mg total RNA using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA, United States). At least three biological replicates were prepared for each sample.