Poly(A) Tail-Length Assay for NOG

A poly(A) tail assay was performed with a Poly(A) Tail-Length Assay Kit (764551KT, Thermo Fisher) according to the protocol. Briefly, poly(A) polymerase was used to add G and I to the 3′ ends of the RNA, and the newly tailed RNA was converted to cDNA by reverse transcription. Then, NOG-specific forward and reverse primers and universal reverse primers were used to generate a product consisting of NOG with a poly(A) tail. The PCR products from METTL3 knockdown DPSCs in differentiated/undifferentiated stages were separated on agarose gels [23 (link)]. The specific primers for NOG were F: 5′-TAACCTGCTATTTATATTCCAGTGCCCTTC-3′ and R: 5′-TGAACTCTATAGCTTCTTCGAGGTCCAA-3′.