GSM8018891: E2D.R1C2 replica 2; Pseudomonas aeruginosa; RNA-Seq
Total RNA was then extracted and purified utilizing the Monarch Total RNA Isolation Kit (New England Biolabs), followed by triple DNase I (Promega) treatment to eliminate any genomic DNA contamination, and subsequently re-purified using a phenol-chloroform extraction method. Firstly, ribosomal RNA was removed by rRNA removal kit, and rRNA free residue was cleaned up by ethanol precipitation. Subsequently, sequencing libraries were generated using the rRNA-depleted RNA and performing procedures as follow. Briefly, after fragmentation, the first strand cDNA was synthesized using random hexamer primers. Then the second strand cDNA was synthesized and dUTPs were replaced with dTTPs in the reaction buffer. The directional library was ready after end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification.