Purification of high-quality micro RNA from the heart tissue

Nóra Faragó, A. Zvara, Z. Varga, P. Ferdinándy, L. Puskás

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Micro RNAs (miRNA) are an abundant class of small RNAs that regulate the stability and translation of cognate mRNAs. MiRNAs are potential diagnostic markers, moreover, they play an essential role in the development of various heart disesases. In case of limited tissue material, such as, e.g. human biopsies, purification of miRNAs with sufficient yield is critical. Reproducible expression analysis of miRNAs is highly dependent on the quality of the RNA, which is often difficult to achieve from fibrous tissue such as the heart. Several companies developed general purification kits for miRNAs, however, none of them are specialized to fibrotic tissues. Here we describe an optimized miRNA purification protocol that results in high miRNA yield as compared to other methods including trizol-based and column-based protocols. By using our improved protocol, miRNA obtained from heart tissue gave more reproducible results in QRT-PCR analysis and obtained more significant calls (172 vs. 118) during DNA microarray analysis when compared to the commercially available kit. In addition to the heart tissue, the present protocol can be applied to other fibrotic tissues, such as lung or skeletal muscle to isolate high-purity miRNA.

Original languageEnglish
Pages (from-to)413-425
Number of pages13
JournalActa Biologica Hungarica
Volume62
Issue number4
DOIs
Publication statusPublished - Dec 1 2011

Fingerprint

MicroRNAs
Purification
purification
RNA
Tissue
tissue
Biopsy
RNA Stability
Protein Biosynthesis
Microarray Analysis
Microarrays
Oligonucleotide Array Sequence Analysis
muscle
Muscle
Skeletal Muscle
DNA
protocol
Polymerase Chain Reaction
Lung
Messenger RNA

Keywords

  • Heart
  • microarray
  • miRNA expression
  • miRNA purification

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Environmental Science(all)
  • Neurology

Cite this

Purification of high-quality micro RNA from the heart tissue. / Faragó, Nóra; Zvara, A.; Varga, Z.; Ferdinándy, P.; Puskás, L.

In: Acta Biologica Hungarica, Vol. 62, No. 4, 01.12.2011, p. 413-425.

Research output: Contribution to journalArticle

@article{bcef795d0b31461f932594e74d0086d8,
title = "Purification of high-quality micro RNA from the heart tissue",
abstract = "Micro RNAs (miRNA) are an abundant class of small RNAs that regulate the stability and translation of cognate mRNAs. MiRNAs are potential diagnostic markers, moreover, they play an essential role in the development of various heart disesases. In case of limited tissue material, such as, e.g. human biopsies, purification of miRNAs with sufficient yield is critical. Reproducible expression analysis of miRNAs is highly dependent on the quality of the RNA, which is often difficult to achieve from fibrous tissue such as the heart. Several companies developed general purification kits for miRNAs, however, none of them are specialized to fibrotic tissues. Here we describe an optimized miRNA purification protocol that results in high miRNA yield as compared to other methods including trizol-based and column-based protocols. By using our improved protocol, miRNA obtained from heart tissue gave more reproducible results in QRT-PCR analysis and obtained more significant calls (172 vs. 118) during DNA microarray analysis when compared to the commercially available kit. In addition to the heart tissue, the present protocol can be applied to other fibrotic tissues, such as lung or skeletal muscle to isolate high-purity miRNA.",
keywords = "Heart, microarray, miRNA expression, miRNA purification",
author = "N{\'o}ra Farag{\'o} and A. Zvara and Z. Varga and P. Ferdin{\'a}ndy and L. Pusk{\'a}s",
year = "2011",
month = "12",
day = "1",
doi = "10.1556/ABiol.62.2011.4.7",
language = "English",
volume = "62",
pages = "413--425",
journal = "Acta Biologica Hungarica",
issn = "0236-5383",
publisher = "Akademiai Kiado",
number = "4",

}

TY - JOUR

T1 - Purification of high-quality micro RNA from the heart tissue

AU - Faragó, Nóra

AU - Zvara, A.

AU - Varga, Z.

AU - Ferdinándy, P.

AU - Puskás, L.

PY - 2011/12/1

Y1 - 2011/12/1

N2 - Micro RNAs (miRNA) are an abundant class of small RNAs that regulate the stability and translation of cognate mRNAs. MiRNAs are potential diagnostic markers, moreover, they play an essential role in the development of various heart disesases. In case of limited tissue material, such as, e.g. human biopsies, purification of miRNAs with sufficient yield is critical. Reproducible expression analysis of miRNAs is highly dependent on the quality of the RNA, which is often difficult to achieve from fibrous tissue such as the heart. Several companies developed general purification kits for miRNAs, however, none of them are specialized to fibrotic tissues. Here we describe an optimized miRNA purification protocol that results in high miRNA yield as compared to other methods including trizol-based and column-based protocols. By using our improved protocol, miRNA obtained from heart tissue gave more reproducible results in QRT-PCR analysis and obtained more significant calls (172 vs. 118) during DNA microarray analysis when compared to the commercially available kit. In addition to the heart tissue, the present protocol can be applied to other fibrotic tissues, such as lung or skeletal muscle to isolate high-purity miRNA.

AB - Micro RNAs (miRNA) are an abundant class of small RNAs that regulate the stability and translation of cognate mRNAs. MiRNAs are potential diagnostic markers, moreover, they play an essential role in the development of various heart disesases. In case of limited tissue material, such as, e.g. human biopsies, purification of miRNAs with sufficient yield is critical. Reproducible expression analysis of miRNAs is highly dependent on the quality of the RNA, which is often difficult to achieve from fibrous tissue such as the heart. Several companies developed general purification kits for miRNAs, however, none of them are specialized to fibrotic tissues. Here we describe an optimized miRNA purification protocol that results in high miRNA yield as compared to other methods including trizol-based and column-based protocols. By using our improved protocol, miRNA obtained from heart tissue gave more reproducible results in QRT-PCR analysis and obtained more significant calls (172 vs. 118) during DNA microarray analysis when compared to the commercially available kit. In addition to the heart tissue, the present protocol can be applied to other fibrotic tissues, such as lung or skeletal muscle to isolate high-purity miRNA.

KW - Heart

KW - microarray

KW - miRNA expression

KW - miRNA purification

UR - http://www.scopus.com/inward/record.url?scp=80053188969&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80053188969&partnerID=8YFLogxK

U2 - 10.1556/ABiol.62.2011.4.7

DO - 10.1556/ABiol.62.2011.4.7

M3 - Article

VL - 62

SP - 413

EP - 425

JO - Acta Biologica Hungarica

JF - Acta Biologica Hungarica

SN - 0236-5383

IS - 4

ER -