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RNA-seq

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...

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Updated: Mar 16, 2026

Retrospective MicroRNA Sequencing: Complementary DNA Library Preparation Protocol Using Formalin-fixed Paraffin-embedded RNA Specimens

Retrospective MicroRNA Sequencing: Complementary DNA Library Preparation Protocol Using Formalin-fixed Paraffin-embedded RNA Specimens

Published on: May 5, 2018

Pre-analytical Best Practices for RNA Sequencing from Small Biopsies and Cytologic Specimens.

Gloria Hopkins Sura¹, Kelly B Engel², Sarah R Greytak³

¹Division of Pathology and Laboratory Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. ghsura@mdanderson.org.

Molecular Diagnosis & Therapy

|March 14, 2026

Summary

RNA sequencing from minimally invasive cancer specimens is crucial for precision oncology. This review outlines best practices for specimen handling to ensure reliable RNA sequencing results, improving patient care and research.

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Last Updated: Mar 16, 2026

Retrospective MicroRNA Sequencing: Complementary DNA Library Preparation Protocol Using Formalin-fixed Paraffin-embedded RNA Specimens

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Published on: May 5, 2018

Sequencing Small Non-coding RNA from Formalin-fixed Tissues and Serum-derived Exosomes from Castration-resistant Prostate Cancer Patients

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Published on: November 19, 2019

Improving Small RNA-seq: Less Bias and Better Detection of 2'-O-Methyl RNAs

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Published on: September 16, 2019

Area of Science:

Molecular Biology
Genomics
Oncology

Background:

RNA sequencing is vital in precision oncology for transcriptome profiling, gene-fusion detection, and biomarker discovery.
Minimally invasive procedures yield specimens ideal for RNA sequencing but present unique pre-analytical challenges.
Pre-analytical factors significantly impact RNA sequencing success, quality, and accuracy, affecting clinical and research outcomes.

Purpose of the Study:

To establish evidence-based pre-analytical best practices for minimally invasive specimens used in RNA sequencing.
To provide guidance for National Cancer Institute researchers and clinicians.
To enhance the reliability and reproducibility of RNA sequencing in oncology.

Main Methods:

Literature review and expert consensus from a National Cancer Institute working group.
Identification and synthesis of best practices for specimen collection, processing, and storage.
Focus on strategies for small tissue and cytology specimens.

Main Results:

Defined strategies for assessing specimen adequacy and RNA quality.
Outlined methods to maximize tumor content and minimize RNA degradation.
Provided practical recommendations for handling minimally invasive specimens.

Conclusions:

Implementing standardized pre-analytical workflows is essential for accurate RNA sequencing.
Best practices improve RNA yield and quality from challenging specimens.
This framework supports robust RNA sequencing in clinical and research oncology settings.