Nucleospin rna mini spin kit

RNA was isolated using Nucleospin® RNA mini spin kit (Macherey-Nagel) according to the manufacturer's instructions. 1 μg of total RNA was reverse transcribed using random primers and MultiScribe™ reverse transcriptase (Applied Biosystems). Real-time PCR analysis was performed by Power SYBR Green (Applied Biosystems) in a MX3005P fluorescence temperature cycler (Stratagene) according to the manufacturer's instructions. The sequences of the primers used for the RT-qPCR analyses were as follows: OGT forward 5′-TGG CTT CAG GAA GGC TAT TG-3′ and reverse 5′-CAA GTC TTT TGG ATG TTC ATA TG-3′; cyclin D1 forward 5′-CAT CTA CAC CGA CAA CTC CAT CC-3′ and reverse 5′-TGT TCA ATG AAA TCG TGC GG-3′; RPLP0 (ribosomal protein large subunit P0) forward 5′-GTG ATG TGC AGC TGA TCA AGA-3′ and reverse 5′-GAT GAC CAG CCC AAA GGA GA-3′.

RNA was isolated using Nucleospin^® RNA mini spin kit (Macherey-Nagel) according to the manufacturer’s instructions. 1µg of total RNA was reverse transcribed using random primers and MultiScribeTM reverse transcriptase (Applied Biosystems). Real-time PCR analysis was performed by Power SYBR Green (Applied Biosystems) in a MX3005P fluorescence temperature cycler (Stratagene) according to the manufacturer’s instructions. Results were normalized with respect to RPLP0 mRNA used as internal control. The primers used for the RT-qPCR analyses are summarized in Supplementary Table S1.