title: Function of Polynucleotide phosphorylase of Pseudomonas aeruginosa
design: Purpose: to study the function of Polynucleotide phosphorylase of Pseudomonas aeruginosa. Bacteria were cultured in LB to an OD600 of 1.0. Bacteria were collected, followed by RNA isolation.  The Transcriptome sequencing and analysis were performed by GENEWIZ (Suzhou, China). Briefly, total RNA of each sample was quantified and qualified by an Agilent 2100 Bioanalyzer (Agilent Technologies). 1 μg total RNA with RIN value above 7 was used for library preparation. Large ribosomal RNA was depleted from bacterial total RNA using RiboMinus Bacteria Module (Invitrogen) and the rRNA-depleted mRNA was then fragmented, and primed with random primers. Pair-end index libraries were constructed according to the manufacturer’s protocol (NEBNext. Ultra. Directional RNA Library Prep Kit for Illumina). The RNA expression analysis was based on the annotations of PAO1 (www.pseudomonas.com). The RSEM software (V 1.2.15) was used to align the input reads against the reference gene with Bowtie2 and expression values were calculated using the FPKM (fragments per kilobase of transcript per million reads) method.