RNA-seq of SCQ1-3M
Firstly, the rRNA was depleted from 1 microgram of total RNA using illumina  MRZB12424 Ribo-Zero rRNA Removal Kit (Bacteria) (Illumina, San Diego, CA, USA ). Then, the first-strand cDNA was synthesized using ProtoScript  Reverse Transcriptase (New England BioLabs, Ipswich, MA, USA) at 25 C for 10 min; 42 C  for 15 min; 70 C for 15 min. The second-strand cDNA was synthesized using  NEBNext Second Strand Synthesis Reaction Buffer and dA TP , dGTP , dCTP , dUTP mix  (New England BioLabs, Ipswich, MA, USA) at 16 C for 1 h. Resulted cDNA was  purified with Agencourt AMPure XP beads (Beckman Coulter, Brea, CA) and end  repaired with NEBNext End Repair Reaction Buffer and Enzyme Mix (New England BioLabs, Ipswich, MA, USA) at 20 C for 30 min; 65 C for 30 min. Sequencing  adapters were ligated using NEBNext Adaptor for Illumina (New England BioLabs, Ipswich, MA, USA) at 20 C for 15 min. The second-strand cDNA was then degraded  using the USER enzyme mix (New England BioLabs, Ipswich, MA, USA) at 37 C for  15 min and the product was purified by Agencourt AMPure XP beads (Beckman Coulter, Brea, CA). Then the sequencing library was constructed using NEBNext Poly(A)  mRNA Magnetic Isolation Module (New England Biolabs, Ipswich, MA, USA).