| Construction Protocol | Un-induced SY5Y-MYCN cells and cells treated with 1ug/ml Doxycycline (Sigma) to induce MYCN overexpression, for 1h, 4h 24h were used. For the time-course treatment start times were staggered so all samples were lysed simultaneously. Total mRNA was extracted using TRI Reagent (Sigma-Aldrich) according to manufacturer’s protocol, and DNA was digested with DNA-free Kit (Applied Biosystems). miRNA libraries (SY5Y-MYCN; un-induced, 1h, 4h and 24h MYCN induction) were prepared using TruSeq small RNA sample preparation kit (Illumina). |
