Small RNA-seq profiling of wild-type Apilactobacillus kunkeei strains in mid-exponential phase using UMI-based libraries
Small RNA-seq profiling of wild-type Apilactobacillus kunkeei strains in mid-exponential phase using UMI-based libraries
Cultures were grown statically in MRS medium supplemented with 5% fructose at 35 °C. Cells were harvested during mid-exponential growth (OD600 0.5–0.6) by centrifugation and immediately stabilized in RNAlater. Samples were stored and transported on dry ice prior to RNA extraction. Cell pellets corresponding to 10 OD units were lysed in TRIzol using a handheld homogenizer. Total RNA was purified using the Ribo-Pure RNA extraction kit (Thermo Fisher), followed by DNase digestion. RNA quality and integrity were assessed prior to library preparation. Small RNA sequencing libraries were prepared at BGI Genomics using a UMI small RNA library preparation kit. RNA fragments of approximately 17–200 nt were size-selected according to the manufacturer's instructions. Adapter ligation and UMI incorporation were performed during library construction. The final libraries were quality-checked and quantified before sequencing.