title: Illumina HiSeq 2000 sequencing; Comparison of RNA-seq of Actinobacillus pleuropneumoniae (WT and (p)ppGpp0) under the absence of branched chain amino acids
design: Comparison of RNA-seq of Actinobacillus pleuropneumoniae (WT and (p)ppGpp0) under the absence of branched chain amino acids
library_construction_protocol: The cells were collected at 4°C, and washed twice with precooled PBS buffer, and the pellets were frozen in liquid nitrogen.  All bacteria were cultured at 37°C.  The WT and (p)ppGpp0 mutant were grown to log-phase in CDM and single BCAA deleted medium The RNeasy kit (Qiagen) and Ribo-Zero™ rRNA Removal Kit (EPICENTRE Biotechnologies) was used to isolate RNA and remove rRNA. Total 20 μg of RNAs for both WT and (p)ppGpp0 mutant were pooled for cDNA library construction. The cDNA libraries were constructed according to Illumina's protocols and sequenced using the Illumina HiSeq 2000 platform.