title: A phenol-free method for isolation of microRNA from biological fluids.
abstract: MicroRNAs (miRNAs) found in biological fluids such as blood and urine have been identified as promising biomarkers for many human disorders, including cancer, cardiopathies, and neurodegenerative diseases. However, circulating miRNAs are either encapsulated into vesicles or found in complexes with proteins and lipoproteins and, thus, require a special approach to their isolation. Acid phenol-chloroform extraction can solve this problem, but it is a labor-intensive procedure that relies heavily on the use of hazardous chemicals. Here we describe a fast and simple phenol-free protocol for miRNA isolation from biofluids. MiRNA is extracted from complexes with biopolymers by a high concentration of guanidine isothiocyanate combined with water/organic composition of solvents. Purification is finished using silica-based spin columns. Comparison of miRNA isolation from blood plasma and urine using the single-phase method and acid phenol-chloroform extraction by means of radioisotope spike-ins and quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) showed similar performance of the two methods.
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine SB RAS (ICBFM SB RAS), Novosibirsk 630090, Russia. Electronic address: ivanzap@niboch.nsc.ru.
authors.forename: Ivan A
authors.initials: IA
authors.lastname: Zaporozhchenko
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine SB RAS (ICBFM SB RAS), Novosibirsk 630090, Russia.
authors.forename: Evgeniy S
authors.initials: ES
authors.lastname: Morozkin
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine SB RAS (ICBFM SB RAS), Novosibirsk 630090, Russia.
authors.forename: Tatyana E
authors.initials: TE
authors.lastname: Skvortsova
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine SB RAS (ICBFM SB RAS), Novosibirsk 630090, Russia.
authors.forename: Olga E
authors.initials: OE
authors.lastname: Bryzgunova
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine SB RAS (ICBFM SB RAS), Novosibirsk 630090, Russia.
authors.forename: Anna A
authors.initials: AA
authors.lastname: Bondar
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine SB RAS (ICBFM SB RAS), Novosibirsk 630090, Russia.
authors.forename: Ekaterina M
authors.initials: EM
authors.lastname: Loseva
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine SB RAS (ICBFM SB RAS), Novosibirsk 630090, Russia.
authors.forename: Valentin V
authors.initials: VV
authors.lastname: Vlassov
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine SB RAS (ICBFM SB RAS), Novosibirsk 630090, Russia.
authors.forename: Pavel P
authors.initials: PP
authors.lastname: Laktionov
mesh_terms: D006146:Guanidines / Q000737:chemistry; D006801:Humans; D017879:Isothiocyanates / Q000737:chemistry; D035683:MicroRNAs / Q000097:blood / Q000302:isolation & purification* / Q000652:urine; D019800:Phenol / Q000737:chemistry; D012822:Silicon Dioxide / Q000737:chemistry; D052616:Solid Phase Extraction / Q000379:methods*; D012997:Solvents
chemical_list: D006146:Guanidines; D017879:Isothiocyanates; D035683:MicroRNAs; D012997:Solvents; C054435:guanidine isothiocyanate; D019800:Phenol; D012822:Silicon Dioxide
keywords: Blood plasma; Circulating miRNA; MiRNA isolation; Phenol free; Single phase; Urine