Ssdna assay kit

Single-indexed cDNA libraries were generated using the SMARTer Stranded RNA-Seq Kit (Clontech Inc.) in accordance with the manufacturer’s instructions. Fluorescent measurement of DNA concentrations in each library was performed using Qubit dsDNA BR and ssDNA assay kit (Thermo Fischer Scientific).

The iodide uptake assay was performed as described previously (29 (link)). In brief, 293T cells were seeded in 12-well plates, which contained sterilized cover slips (WHB Biotech), incubated at 37°C for 24 h, and transiently transfected alone or co-transfected with WT or mutant pEGFP-N2 plasmids. 48 h after transfection, the 293T cells were washed once in serum-free DMEM medium and incubated for 1 h in 1 ml serum-free medium containing ¹³¹I at 5 KBq/ml as the only source of iodide. For the inhibition of NIS-mediated uptake, NaClO₄ (final concentration 1 mM) was included in parallel incubations (30 (link)). The cells were washed briefly in Hank’s Balanced Salt Solution (HBSS) buffer and then incubated with 1 ml HBSS for 5 min. The cells were solubilized by the addition of 1 ml 1 M NaOH, and the radioactivity was measured using a γ counter (GC1200, Anhui, China). DNA was determined by the Qubit Fluorometer using ssDNA Assay Kit (Q10212, Thermofisher). Each value represents the mean ± SD of picomoles ¹³¹I per microgram of DNA of three independent experiments done in triplicate.