The total RNA was prepared using the RNA Extraction Kit (QIAGEN, Germany) and rRNA was removed using Ribo-Zero rRNA Removal Kit (Epicentre, United States), following the manufacturer’s protocol. Sequencing libraries were generated using the NEBNext Ultra Directional RNA Library Prep Kit for ...
RNA-Seq Analysis To compare how TCS exposure changed bacterial gene expression, we conducted RNA-Seq analysis for the parental (Kp85anc) and one representative evolvedK. pneumoniaestrain (d10-2) in the absence and presence of TCS. Three replicates were conducted for each condition, and the dat...
RNA isolation and real-time quantitative reverse transcriptase (qRT)-PCR Total RNA was isolated from cells using RNAeasy Mini kit (Qiagen, Georgetown, MD) according to manufacturer’s instructions. RevertAid first strand cDNA synthesis kit (Fermentas life sciences, Cromwell Park, MD) was used to ...
16S ribosomal RNA high-throughput sequencing was used to identify, classify, and predict the functions of the bacterial communities in the eggs and ovaries ofBactrocera cucurbitae(Coquillett) (Diptera: Tephritidae), which is a pest that infests a variety of cucurbit fruits at different developmental...
RNA was isolated with the Agilent Total RNA Isolation Mini Kit following the detailed protocol in Appendix A, or by a silica-based method (QIAGEN RNeasy and RNase-free DNase Set, QIAGEN, Valencia, CA). RNA quality was determined via the Prokaryotic Total RNA Nano asasy from the RNA 6000 ...
For microaerobic free-living conditions, 50 ml of bacterial cell cultures were collected and total RNA isolated using a RNeasy Mini Kit (QIAGEN, Hilden, Germany). RNA concentration was determined by measuring the absorbance at 260 nm. The integrity of RNA was determined by running samples on ...
Methods and Results Human milk DNA was extracted using the following methods: (i) Qiagen MagAttract Microbial DNA Isolation Kit (kit QM), (ii) Norgen Milk Bacterial DNA Isolation Kit (kit NM), (iii) Qiagen MagAttract Microbiome DNA/RNA Isolation Kit (kit MM) and (iv) TRIzol LS Reagent (...
RNA extraction and quantitative real-time PCR analysis Total RNA was extracted using a RNeasy Plus Mini Kit (Qiagen) following the manufacturer’s instructions. Briefly, cells were washed with PBS in a 6-well plate. To avoid any gene expression changes during trypsinisation, cells were lysed ...
Isolation of RNA. Total RNA was extracted from bacterial pellets using the RNeasy Mini kit (Qiagen) according to the manufacturer's instruction. Potential traces of DNA were removed by digestion with DNase I (Turbo DNA-free, Ambion), according to the manufacturer's instructions. RNA ...
RNA was isolated from the samples using a Promega SV total RNA isolation kit with the RNA quality being verified by an Agilent Bioanalyser using the RNA Nano kit. RT-PCR was performed with the Qiagen One-step RT-PCR kit with the following reaction conditions: One cycle 50 °C for 30...