RNA splicing dysregualtion in the pathogenesis of chronic lymphocytic leukemia: Sample N_42
PolyA mRNA was heated at 85°C for 5 minutes and chilled on ice to denature RNA secondary structure. Then, 100ng denatured polyA mRNA was digested with 10 Unit of MazF enzyme (#2415A, TaKaRa) in MazF reaction buffer at 37°C for 30 min. The digested RNA was purified by RNA Clean & Concentrator-5 Kit (Zymo Research) and ends were repaired by T4 Polynucleotide Kinase (#M0201S, NEB). The sequencing library was constructed by using the VAHTS Small RNA Library Prep Kit (Kit #NR801, Vazyme Biotech) with 10 ng of fragmented polyA RNA and was amplified by PCR for 15 cycles.