Open AccessIsolation of DNA-free RNA from human bone marrow mononuclear cells: comparison of laboratory methods
Author(s) -
Tawakalitou Alabi,
Sweta B. Patel,
Smita Bhatia,
Julie Wolfson,
Purnima Singh
Publication year - 2020
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2019-0093
Subject(s) - rna , rna extraction , peripheral blood mononuclear cell , bone marrow , biology , dna , gene expression , microbiology and biotechnology , computational biology , polymerase chain reaction , gene , immunology , genetics , in vitro
RNA quality (purity and integrity) and quantity are of critical importance to ensure reliable gene expression analysis, reproducibility of RNA sequencing and microarray data and validation by RT-PCR. Currently available methods for isolating RNA either are labor intensive (requiring the use of toxic organic solvents and separate DNase treatment) or require automation (with extensive setup and startup costs). To optimize both the quality and quantity of RNA from bone marrow, we recommend stabilization and storage of bone marrow mononuclear cells in RNAprotect ® Cell Reagent, followed by extraction using the RNeasy ® Protect Cell Mini Kit (Qiagen, Hilden, Germany). This method achieves optimal quantity and high-quality RNA for sequencing and RT-PCR while remaining efficient and cost effective.