MagExtractor -RNA

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	Total RNA was collected from the cultured COV434 cells using the MagExtractorTM RNA kit (Toyobo). The RNA was reverse transcribed using the iScript cDNA Synthesis kit (Bio-Rad), according to the manufacturer’s protocol.

Protocol tips
Total RNA was collected from the cultured COV434 cells using the MagExtractorTM RNA kit (Toyobo). The RNA was reverse transcribed using the iScript cDNA Synthesis kit (Bio-Rad), according to the manufacturer’s protocol.

Publication protocol

Total RNA was collected from the cultured COV434 cells using the MagExtractorTM RNA kit (Toyobo). The RNA was reverse transcribed using the iScript cDNA Synthesis kit (Bio-Rad), according to the manufacturer’s protocol. Aliquots of cDNA were amplified in iQ SYBR Green Supermix (Bio-Rad) using a CFX96 real-time PCR detection system (Bio-Rad). The temperature profiles used for PCR comprised denaturation for 30 sec at 95°C, annealing for 30 sec at 60°C and elongation for 1 min at 72°C, with a total of 40 cycles. The target SF-1 mRNA was normalized against the housekeeping enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA as an internal control. The results are expressed as the ratio relative to the control. The forward and reverse oligonucleotide primers were designed on the basis of published cDNA sequences. The primer sequences were as follows: GAPDH forward 5'-GAGTC AACGGATTTGGTCGT-3'; GAPDH reverse 5'-TTGATT TTGGAGGGATCTCG-3'; SF-1 forward 5'-GCTACCCA GAGCCCTTCTCT-3'; SF-1 reverse 5'-TCCAGCTCCTTG AAGACCAT-3'. The results are expressed as the mean±SD of triplicate samples (n=3).

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Manufacturer protocol

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