lamdaN-nrde-2_nrde-4_reporter rep2
Trizol extracted total RNA was subjected to mirVana enrichment for small RNAs. Homemade 5' polyphosphatase PIR-1 was used to remove the g and b phosphates of 22G-RNAs. Adaptors of 3' and 5' were ligated to small RNA with T4 RNA ligase 2 (NEB), and T4 RNA ligase 1(NEB) sequentially. After reverse transcription using Superscript III (Thermo Fischer Scientific), the cDNA was amplified by PCR. Raw reads need to be trimmed by TGGAATTCTC at 3' and 15 nt at 5' (UMI).