Trizol max bacterial enhancement kit

Biofilms of H. pylori SS1 were grown in 6-well plates (Costar) in BB2 as described above. After 3 days of incubation, medium containing nonattached planktonic bacteria (the planktonic fraction) was removed by pipetting, and the cells were harvested by centrifugation and washed twice with PBS. The attached bacteria, representing the biofilm fraction, were washed twice with PBS to remove any remaining planktonic cells. Attached cells were scrapped off the plate using a cell scraper. Both planktonic and biofilm fractions were subject to total RNA extraction using the TRIzol Max bacterial enhancement kit (Ambion, Life Technology, Carlsbad, CA) as described by the manufacturer. RNA was further purified and concentrated using an RNeasy kit (Qiagen). rRNA was removed using the RiboZero magnetic kit (Illumina). Sequencing libraries were generated using NEBNext Ultra Directional RNA library prep kit for Illumina (NEB, USA). cDNA library quality and amount were verified using the Agilent Bioanalyzer 2100 system (Agilent Technologies, CA) and then sequenced using Illumina NextSeq Mid-Output (University of California Davis Genome Center).

Total RNA from H. pylori in Trizol Max was extracted using the Trizol Max Bacterial Enhancement Kit (Ambion, Life Technology, Carlsbad, CA, USA) as described by the manufacturer. RNA was further purified and concentrated using an RNAeasy Kit (Qiagen). rRNA was removed using RiboZero magnetic kit (Illumina). Sequencing libraries were generated using NEBNext Ultra^TM Directional RNA library Prep Kit for Illumina (NEB, USA). Complementary DNA (cDNA) library quality and amount were verified using Agilent Bioanalyzer 2100 system (Agilent technologies, CA, USA) and then sequenced using Illumina NextSeq Mid-Output (UC Davis Genome Center).