Table 3.
Comparative studies of different miRNA isolation methods.
| The Performance of Different Isolation Methods in Terms of Yield, Purity and miRNAs Detection Level | miRNAs & Detection Method | Sample Type | References |
|---|---|---|---|
| Total RNA isolation kit < miRNeasy kit, the miRVana PARIS kit | miRCURY LNA real-time PCR panel | serum | [134] |
| TRIzol-based miRNA isolation<The miRNeasy Mini kit | qRT-PCR miR-92a, -126 | plasma, serum | [135] |
| miRvana microRNA Isolation kit, QIAamp Circulating Nucleic Acid Kit > miRNeasy Serum/Plasma Kit | qRT-PCR miR-21, -191, syntheticath-miR-159a | plasma | [136] |
| miRNeasy Serum/Plasma kit, MagnaZol™ cfRNA Isolation Reagent | NGS | plasma | [137] |
| miRCURY Cell and Plant Kit without glycogen < TRIzol LS (LT) followed by mirVana column clean-up with glycogen/without glycogen< miRCURY Cell and Plant Kit with glycogen < mirVana kit with glycogen/without glycogen miRCURY Cell and Plant Kit with glycogen < miRCURY Biofluids kit with glycogen/without glycogen miRNeasy Serum/Plasma system slightly < miRCURY Biofluids kit |
qRT-PCR miR-16, -21, cel-miR-39 | plasma | [131] |
| TRIzol LS (LT); miRNA purification kit < mirVana PARIS kit | qRT-PCR cel-miR-39 | plasma | [138] |
| TRIzol LS (LT); miRNeasy mini kit; mirVana PARIS kit; miRNA purification kit | qRT-PCR miR-21 | ||
| mirVana PARIS, Trizol LS (LT) with mirVana, miRCURY < miRNeasy, < miRNeasy with RNeasy MinElute Cleanup Kit | Bioanalyser | urinary exosomes | [106] |
| mirVana PARIS (with addition of an additional organic extraction step), < miRNeasy kit | qRT-PCR | plasma serum |
[139] |
| comparable results: miRNeasy mini kit, Plasma/Serum Circulating and Exosomal RNA Purification Kit = NucleoSpin miRNAs Plasma kit | Microarray | peripheral blood mononuclear cells | [127] |
| MirVana PARIS kit, TRIzol-LS (LT) < miRNeasy Serum/Plasma Kit | fresh plasma | ||
| TRIzol-LS (LT) < mirVana PARIS kit, TRIzol-LS following mirVana kit < miRNeasy Serum/Plasma kit | frozen plasma | ||
| comparable results in terms of Cq values: miRCURY RNA Isolation Kits, Plasma/Serum Circulating and Exosomal RNA Purification Mini Kit, NucleoSpin miRNA Plasma, miRNeasy Serum/Plasma Kit, Direct-zol RNA MiniPrep |
miRCURY microRNA QC PCR Panel (Exiqon) miR-18a, -21, -29a | plasma | [140] |
| TRIzol-LS (LT) < miRNeasy, mirVana miRNA Isolation Kit < TRIzol-LS (LT) combined with miRNeasy < miRCURY RNA Isolation Kit < SeraMir exoRNA columns | RT-qPCR, mir-16 and let-7i |
UE purified by UF or from peripheral blood mononuclear cells | [141] |
| PAXgene and Tempus whole blood RNA preservation tubes Qiagen (PAXgene Blood miRNA Kit), Life Technologies (MagMAX for Stabilized Blood Tubes RNA Isolation Kit), Norgen Biotek (Norgen Preserved Blood RNA Purification Kit I and Kit II) and 2 (semi-) automated protocols on the QIAsymphony (Qiagen) and MagMAX Express-96 Magnetic Particle Processor (Life Technologies) Norgen Kit II with PAXgene < Norgen Kit I with Tempus Tubes |
qRT-PCR miR-1, -16, -30b, -133a |
blood | [140] |
| QIAGEN Exo kit < UC fraction using the miRNeasy kits and the NucleoSpin kit | miRCURY microRNA QC PCR Panel (Exiqon) miR-103, -191, -23a, -451, -18a, -21, -29a |
EVs | [142] |
| Similar Cq values: miRNeasy Mini Kit, miRNeasy Serum/Plasma kit, exoRNeasy Serum/Plasma Starter Kit, Plasma/Serum RNA Purification Mini Kit, Direct-zol™ RNA MiniPrep, NucleoSpin miRNA Plasma | plasma | ||
| miRNeasy slightly better than miRCURY | RT-qPCR, mir-16, -106a, -222, -223 | plasma exosomes | [133] |
| miRCUR < miRNeasy miRCURY biofluids = miRNeasy kits |
RT-qPCR, mir-30c-2-3p, -106a, -204, -222, -141 | plasma | |
| miRCURY < miRNeasy | urinary exosome | ||
| miRNeasy Serum/Plasma kit, Plasma/Serum Circulating RNA Purification Kit < miRCURY RNA Isolation Kit—Biofluids; mirVana PARIS Kit; NucleoSpin miRNA Plasma | TaqMan miRNA PCR system (Applied Biosystems, California, USA), miRCURY LNA microRNA PCR system (Exiqon) and miScript miRNA PCR system (Qiagen), miR-16, -141, -135a | plasma | |
| PureLink commercial column extraction kit < TiO2 nanofibers | qRT-PCR miR-21, -191, spiked cel-miR-54 | MDA-MB-231 breast cancer cells | [133] |
| Phenol and column-based procedure and a column-based procedure, in the presence or absence of two RNA carriers (yeast RNA and MS2 RNA); others carriers and their absence < yeast RNA | NGS | plasma | [143] |
| ExoRNeasy Serum/Plasma Starter Kit (Cat. 77023), Plasma/Serum RNA Purification Mini Kit, Direct-zol™ RNA MiniPrep < miRNeasy kits (Mini and Serum/Plasma kits) and NucleoSpin kit | Bioanalyzer, qRT-PCR miR-122, -21-1, -30d, -451a |
plasma | [144] |
| Trizol LS reagent isolation method < miRNeasy mini kit by 35% | qRT-PCR RNU6, miR-145, -20a | serum | [145] |
| Trizol LS < miRCURY < miRNeasy kit, with some peculiarities concerning different sample volumes | NGS, qRT-PCR miR-106a, -222, -16, -223 | plasma and circulating exosomes | [146] |
| mirVana and the Plasma/Serum RNA Purification Mini < miRNeasy Serum/Plasma < miRCURY Biofluids and the miRNeasy Advanced Serum/Plasma | NGS, qRT-PCR let-7a-5p, miR-150-5p, -16-5p, -122–5p, -21-5p, -191-5p, cel-miR-39-3p | plasma cf-miRNA | [122] |
| exoRNeasy Serum/Plasma Exosome slightly <Ultracentrifugation + miRNeasy kit | plasma EVmiRNA | ||
| Circulating Nucleic Acid Kit < TRIzol LS (TF); miRNEasy; RNA extraction kit and the MiRCURY RNA Isolation Kit—intermediate | NGS | plasma | [147] |
| miRNeasy Serum/Plasma kit (Qiagen, Germany), miRVANA miRNA Isolation Kit (Ambion, Austin, TX, USA) and TRIzol LS (TF) < Urine microRNA purification kit | qRT-qPCR miR-16, -10a-5p, -196a-5p) and exogenous cel-miR-39 | CSF samples | [148] |
| Quick-cfRNA Serum and plasma; Isolate II miRNA kit; PureLink RNA mini kit; Monarch total RNA miniprep kit < miRNeasy Serum/Plasma Advanced kit < miRNeasy Serum/Plasma kit |
qRT-PCR miR-19b, -92a, -93, -103, -144, -345-3p, -486, -494-3p, -1306 | ovine plasma, fresh and frozen | [70] |
<, >—better or worth in terms of yield, purity and miRNAs detection level. Total RNA isolation kit, (Norgen, Thorold, Ontario, Canada);Plasma/Serum Circulating and Exosomal RNA Purification Mini Kit, (Norgen, Thorold, Ontario, Canada);Urine microRNA purification kit, (Norgen, Thorold, Ontario, Canada);MiRNeasy kit, (Qiagen, Hilden, Germany);QIAamp® Circulating Nucleic Acid Kit, (Qiagen, Hilden, Germany);ExoRNeasy Serum/Plasma Starter Kit, (Qiagen, Hilden, Germany);MiRNeasy Serum/Plasma Kit, (Qiagen, Hilden, Germany);MiRNeasy with RNeasy MinElute Cleanup Kit, (Qiagen, Hilden, Germany);QiaSymphony RNA extraction kit, (Qiagen, Hilden, Germany);MiRVana PARIS (Ambion, Life Technologies, Austin, TX, USA); MiRvana microRNA Isolation kit (Ambion, Life Technologies, Austin, TX, USA ); Trizol LS reagent (LT) (Ambion, Life Technologies, Carlsbad, CA, USA); MiRCURY RNA isolation kit Biofluids (Exiqon, Vedbaek, Denmark); MiRCURY Cell and Plant Kit (Exiqon, Vedbaek, Denmark); MiRCURY Biofluids kit (Exiqon, Vedbaek, Denmark); MagnaZol™ cfRNA Isolation Reagent (Bioo Scientific, Austin, TX, USA ); ThermoFisher Scientific Ambion TRIzol LS Reagent (TF) (Thermo Fisher Scientific, Third Avenue Waltham, MA, USA); PureLink commercial column extraction kit (Thermo Fisher Scientific, Third Avenue Waltham, MA, USA ); NucleoSpin miRNA Plasma kit (Macherey–Nagel, Düren, Germany); Direct-zol RNA MiniPrep (Zymo Research, Irvine, CA, USA); Quick-cfRNA Serum and plasma (Zymo Research, Irvine, CA, USA); SeraMirTM exoRNA columns (System Biosciences, Palo Alto, CA, USA); PureLink RNA mini kit (Bioline, Memphis, TN, USA); Monarch total RNA miniprep kit (NEB, Hitchin, UK).