Platinum sybr green kit

Total RNA was isolated from cells using NucleoSpin kits (Macherey-Nagel, Bethlehem, PA, USA). One microgram of total RNA was used for q-PCR analysis. cDNA was synthesized using ReverTra Ace cDNA synthesis kits (TOYOBO, Nipro, Osaka, Japan), and 1 μL of the cDNA synthesis reaction mixture was used with a Platinum SYBR Green kit from Qiagen (Hilden, Germany). q-PCR was performed using a Rotor-Gene Q Real-time PCR machine (Qiagen, Hilden, Germany). The oligonucleotides used in PCR were as follows: Kdm2b forward, 5′-AGCAGCTAAAACCTGGCAAA-3′, and reverse, 5′-GTGAGCTGGAACGTGACTGA-3′; Acta1 forward, 5′-GACCTCACTGACTACCTGATGAAA-3′, and reverse, 5′-CAGACTCCATACCGATAAAGGAAG-3′; myogenin forward, 5′-AGTACATTGAGCGCCTACAG-3′, and reverse, 5′-ACCCACCCTGACAGACAATC-3′; Mck forward, 5′-AGCAGCTCATTGATGACCAC-3′, and reverse, 5′-TCAAACTTGGGGTGCTTGCT-3′; Myod forward, 5’-TGCTCTGATGGCATGATGGA-3’, and reverse, 5’-CACTATGCTGGACAGGCAGT-3’; Set7 forward, 5’-TGAGGATGGAGGTGTTCTCC-3’, and reverse, 5’-TCTCCCGTCATCTCTCCATC-3’; and beta-actin forward, 5′-CACGATGGAGGGGCCGGACTCATC-3′, and reverse, 5′-TAAAGACCTCTATGCCAACACAGT-3′.