GSM6374052: P_Pa3; Pseudomonas aeruginosa; RNA-Seq
The bacterial collection was added with phenol  and shaken to be mixed. The centrifuge tube was incubated at 65℃ with an oscillating metal bath at maximum speed for 30–60 minutes, cooled on ice for 5 minutes, then centrifuged at 4℃ for 10 minutes. The upper water phage was absorbed and added with 1/2 volume of TRK-1002 lysate. The concentration and purity of RNA were measured using an Agilent 2100 bioanalyzer.An RNA integrity number (RIN) >7.0 can be used to construct a cDNA library Ribosomal RNA were removed from the total RNA by using a Ribo-Zero Magnetic Kit , and then the library was generated using an Illumina Truseq RNA Sample Preparation Kit according to the manufacturer's recommendation. After digested into 200 nt fragments, double-stranded cDNAs were synthesized using uracil-N-glycosylase (UNG) to a cDNA cluster. The cDNAs were used to create the RNA-seq library by following the standard Illumina protocol.