title: HCC_dual_small_18
design: Resected tumor sample was collected from a 46-year-old male patient and transported in DMEM (GIBCO) on ice immediately after surgical. Tumor pieces were digested with accumax to obtain single cell suspension. Single cells were picked up and lysed with 200 ng of the carrier RNA mixture (containing 100 ng 5'-phosphorylated small carrier RNA mimic). The poly-A RNAs were selected using a NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490), and the small RNAs remained in the supernatant after poly-A selection. The small RNA sequencing library was constructed following the manufacturer's protocol using a NEBNext kit (NEB #E7300). After PCR amplification (15 cycles), the cDNA fragment from the RNA carrier was removed by Not I digestion. After Not I digestion, the library DNA was purified for deep sequencing.