RNA Sequencing Analysis of Salivary Extracellular RNA

Methods Mol Biol. 2017:1537:17-36. doi: 10.1007/978-1-4939-6685-1_2.

Abstract

Salivary biomarkers for disease detection, diagnostic and prognostic assessments have become increasingly well established in recent years. In this chapter we explain the current leading technology that has been used to characterize salivary non-coding RNAs (ncRNAs) from the extracellular RNA (exRNA) fraction: HiSeq from Illumina® platform for RNA sequencing. Therefore, the chapter is divided into two main sections regarding the type of the library constructed (small and long ncRNA libraries), from saliva collection, RNA extraction and quantification to cDNA library generation and corresponding QCs. Using these invaluable technical tools, one can identify thousands of ncRNA species in saliva. These methods indicate that salivary exRNA provides an efficient medium for biomarker discovery of oral and systemic diseases.

Keywords: Biomarkers; RNA sequencing; Saliva; Small and long ncRNA profiling; exRNAs.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Biomarkers
  • DNA Barcoding, Taxonomic
  • Gene Library
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Liquid Biopsy
  • RNA / genetics*
  • RNA / isolation & purification
  • RNA, Long Noncoding
  • RNA, Small Untranslated
  • Saliva / metabolism*
  • Sequence Analysis, RNA*

Substances

  • Biomarkers
  • RNA, Long Noncoding
  • RNA, Small Untranslated
  • RNA