Qpcr detection kit

The human HeLa (ATCC CCL-2) and SH-SY5Y (ATCC CRL-2266) cell lines were maintained in Dulbecco’s Modified Eagle Medium (DMEM) or minimum essential medium α (MEM-α) medium, respectively. Media were supplemented with 10% (v/v) Fetal Bovine Serum (FBS). Both cell lines were grown under a highly humidified atmosphere of 95% air with 5% CO₂ at 37 °C. The HeLa hnRNPDL KO cell line was generated as described elsewhere^{3 (link)}. HeLa cells have been authenticated within the last 3 years by STR analysis. The HeLa and SH-SY5Y cell lines were regularly tested for the presence of mycoplasma using qPCR Detection Kit (SIGMA), and both cell lines are mycoplasma negative.
For recombinant protein expression, the genes encoding the three isoforms of hnRNPDL (hnRNPDL-1, hnRNPDL-2 and hnRNPDL-3) were inserted into pETite (Lucigen corporation) vector with a His-SUMO N-terminal tag. For subcellular localization experiments, the same genes were cloned into pEGFP-C3 (Clontech). In all the cases, the correctness of the DNA sequence was verified by sequencing.

A human melanogenic cell line (A375) and a human immortal keratinocyte line (HaCaT) were obtained from KeyGen Biotech (Nanjing, China) in 2013 and authenticated by short tandem repeat genotyping. Mycoplasma was analyzed using a Mycoplasma quantitative polymerase chain reaction (qPCR) detection kit (Sigma) before experiment. Cells were grown in DMEM supplemented with 10% FBS and maintained at 37 °C in a humidified atmosphere containing 5% CO₂.