Optimization of mRNA extraction from human nasal mucosa biopsies for gene expression profile analysis by qRT-PCR

Publikation: Beitrag in FachzeitschriftForschungsartikelBeigetragenBegutachtung

Beitragende

  • A. U. Bräuer - , Carl von Ossietzky Universität Oldenburg (Autor:in)
  • J. Sevecke-Rave - , Carl von Ossietzky Universität Oldenburg (Autor:in)
  • A. Gläser - , Carl von Ossietzky Universität Oldenburg (Autor:in)
  • P. Nahrath - , Medizinische Fakultät Carl Gustav Carus Dresden (Autor:in)
  • T. Hummel - , Klinik und Poliklinik für Hals-Nasen-Ohrenheilkunde (Autor:in)
  • M. Witt - , Universität Rostock (Autor:in)

Abstract

Quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) is the gold-standard method for analyzing modifications in gene expression in cells and tissues. However, large quantities of high-quality RNA samples are needed for analyzing the expression of multiple genes from one human tissue sample. Here, we provide an optimized protocol for extracting large amounts of RNA from human nasal mucosal biopsies. The quality and quantity of samples were sufficient for qRT-PCR analyses of the expressions of various genes, in duplicate. In contrast to other protocols, we optimized RNA isolation to increase the amount from nasal biopsy samples for analyses of multiple genes. In most previous publications, expressions of only one or a few genes, including housekeeping genes, were analyzed because the amount of biological material was small. We were able to improve our protocol with respect to the yield and quality of RNA. This is likely to produce better results from molecular analyses of very small biopsy samples of human nasal mucosa.

Details

OriginalspracheEnglisch
Seiten (von - bis)1001-1006
Seitenumfang6
FachzeitschriftClinical Anatomy
Jahrgang36 (2023)
Ausgabenummer7
PublikationsstatusVeröffentlicht - 19 Juni 2023
Peer-Review-StatusJa

Externe IDs

PubMed 37337364
ORCID /0000-0001-9713-0183/work/146645739

Schlagworte

Ziele für nachhaltige Entwicklung

ASJC Scopus Sachgebiete

Schlagwörter

  • GAPDH expression, cDNA synthesis, human tissue, ß-actin expression, Nasal Mucosa/chemistry, RNA, Messenger/isolation & purification, Humans, Gene Expression Profiling, Analytic Sample Preparation Methods, Reverse Transcriptase Polymerase Chain Reaction, Real-Time Polymerase Chain Reaction

Bibliotheksschlagworte