RNA was isolated essentially as described (Collart and Oliviero 2001 ) followed by purification using the RNEasy Mini Kit (QIAGEN, 74,104) and DNase I digestion (Invitrogen, 18068-015). Quantitative RT-PCR was carried out using the Superscript III Platinum SYBR Green One-Step qRT-PCR kit (Invitrogen, 11736-059). RNA samples were normalized relative to the BUD6 mRNA levels as previously described (Holstein et al. 2014 (link)).
Free full text: Click here