Illumina NovaSeq 6000 sequencing; RNAseq analysis of samples from oral cancer and oral leukoplakias patients
RNAseq analysis of samples from oral cancer and oral leukoplakias patients
Collection of 37 samples from patients visited and treated at the Stomatology and Maxillofacial Surgery Department of the General University Hospital of Valencia. For all patients, two representative biopsies were taken from the same area of the lesions, including epithelium and the underlying connective tissue. One of each pair of specimens was analyzed with the routine histopathological methods to ensure that the observed lesions meet the histopathological criteria to establish the diagnosis together with the clinical data of each patient. The other sample was used for the RNAseq analysis. In this case, tissue samples were immediately frozen at -80 °C until their analysis. Samples were extracted using column-based DNA extraction method (E.Z.N.A. DNA kit and DNeasy Blood & Tissue Kit; Qiagen, Hilden, Germany) following manufacturer's instructions. cDNA synthesis and library generation were performed using the Illumina TruSeq RNA Library Prep Kit with PolyA selection for ribo depletion, following the manufacturer's instructions.