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Methods Mol Biol. 2017;1537:17-36.

RNA Sequencing Analysis of Salivary Extracellular RNA.

Majem B1, Li F2, Sun J3, Wong DT4,5,6,7.

Author information

1
Biomedical Research Unit in Gynecology, Laboratory 209, Collserola Building, Vall Hebron Research Institute (VHIR) and University Hospital, University Autonoma of Barcelona (UAB), Pg. Vall Hebron 119-129, 08035, Barcelona, Spain.
2
Division of Oral Biology and Oral Medicine, School of Dentistry, University of California Los Angeles (UCLA), 10833 Le Conte Avenue, Los Angeles, CA, 90095, USA.
3
Medical School of Shenzhen University, Shenzhen, Guangdong, China.
4
Division of Oral Biology and Oral Medicine, School of Dentistry, University of California Los Angeles (UCLA), 10833 Le Conte Avenue, Los Angeles, CA, 90095, USA. dtww@ucla.edu.
5
Johnson Comprehensive Cancer Center, University of California Los Angeles (UCLA), Los Angeles, CA, USA. dtww@ucla.edu.
6
Molecular Biology Institute, University of California Los Angeles (UCLA), Los Angeles, CA, USA. dtww@ucla.edu.
7
Head & Neck Surgery/Otolaryngology, Henry Samuel School of Engineering and Applied Science, University of California Los Angeles (UCLA), Los Angeles, CA, 90095-1668, USA. dtww@ucla.edu.

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

PMID:
27924586
PMCID:
PMC5550740
DOI:
10.1007/978-1-4939-6685-1_2
[Indexed for MEDLINE]
Free PMC Article

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