Improved denaturation of small RNA duplexes and it's application for northern blotting

C. Jake Harris; David C. Baulcombe; Attila Molnar

doi:10.1007/978-1-4939-6866-4_1

Improved denaturation of small RNA duplexes and it's application for northern blotting

C. Jake Harris^*, David C. Baulcombe, Attila Molnar

^*Corresponding author for this work

School of Biological Sciences

Research output: Chapter in Book/Report/Conference proceeding › Chapter

Abstract / Description of output

Small RNAs (sRNAs) are short (18-30 nucleotide) noncoding RNA molecules, which control gene expression and pathogen response in eukaryotes. They are associated with and guide nucleases to target nucleic acids by nucleotide base pairing. We found that current techniques for small RNA detection are adversely affected by the presence of complementary RNA. Thus we established FDF-PAGE (fully denaturing formaldehyde polyacrylamide gel electrophoresis), which dramatically improves denaturation efficiency and subsequently the detection of sequestered sRNAs.

Original language	English
Title of host publication	Microrna detection and target identification
Subtitle of host publication	Methods and protocols
Editors	T Dalmay
Publisher	HUMANA PRESS INC
Pages	1-6
Number of pages	6
ISBN (Electronic)	978-1-4939-6866-4
ISBN (Print)	978-1-4939-6864-0
DOIs	https://doi.org/10.1007/978-1-4939-6866-4_1
Publication status	E-pub ahead of print - 25 Apr 2017

Publication series

Name	Methods in Molecular Biology
Publisher	HUMANA PRESS INC
Volume	1580
ISSN (Print)	1064-3745

Keywords / Materials (for Non-textual outputs)

small RNA
sequestration
polyacrylamide gel electrophoresis
denaturation
improved detection

Access to Document

10.1007/978-1-4939-6866-4_1

Cite this

Harris, C. J., Baulcombe, D. C., & Molnar, A. (2017). Improved denaturation of small RNA duplexes and it's application for northern blotting. In T. Dalmay (Ed.), Microrna detection and target identification: Methods and protocols (pp. 1-6). (Methods in Molecular Biology; Vol. 1580). HUMANA PRESS INC. Advance online publication. https://doi.org/10.1007/978-1-4939-6866-4_1

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title = "Improved denaturation of small RNA duplexes and it's application for northern blotting",

abstract = "Small RNAs (sRNAs) are short (18-30 nucleotide) noncoding RNA molecules, which control gene expression and pathogen response in eukaryotes. They are associated with and guide nucleases to target nucleic acids by nucleotide base pairing. We found that current techniques for small RNA detection are adversely affected by the presence of complementary RNA. Thus we established FDF-PAGE (fully denaturing formaldehyde polyacrylamide gel electrophoresis), which dramatically improves denaturation efficiency and subsequently the detection of sequestered sRNAs.",

keywords = "small RNA, sequestration, polyacrylamide gel electrophoresis, denaturation, improved detection",

author = "Harris, {C. Jake} and Baulcombe, {David C.} and Attila Molnar",

year = "2017",

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doi = "10.1007/978-1-4939-6866-4_1",

language = "English",

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series = "Methods in Molecular Biology",

publisher = "HUMANA PRESS INC",

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Improved denaturation of small RNA duplexes and it's application for northern blotting. / Harris, C. Jake; Baulcombe, David C.; Molnar, Attila.
Microrna detection and target identification: Methods and protocols. ed. / T Dalmay. HUMANA PRESS INC, 2017. p. 1-6 (Methods in Molecular Biology; Vol. 1580).

Research output: Chapter in Book/Report/Conference proceeding › Chapter

TY - CHAP

T1 - Improved denaturation of small RNA duplexes and it's application for northern blotting

AU - Harris, C. Jake

AU - Baulcombe, David C.

AU - Molnar, Attila

PY - 2017/4/25

Y1 - 2017/4/25

N2 - Small RNAs (sRNAs) are short (18-30 nucleotide) noncoding RNA molecules, which control gene expression and pathogen response in eukaryotes. They are associated with and guide nucleases to target nucleic acids by nucleotide base pairing. We found that current techniques for small RNA detection are adversely affected by the presence of complementary RNA. Thus we established FDF-PAGE (fully denaturing formaldehyde polyacrylamide gel electrophoresis), which dramatically improves denaturation efficiency and subsequently the detection of sequestered sRNAs.

AB - Small RNAs (sRNAs) are short (18-30 nucleotide) noncoding RNA molecules, which control gene expression and pathogen response in eukaryotes. They are associated with and guide nucleases to target nucleic acids by nucleotide base pairing. We found that current techniques for small RNA detection are adversely affected by the presence of complementary RNA. Thus we established FDF-PAGE (fully denaturing formaldehyde polyacrylamide gel electrophoresis), which dramatically improves denaturation efficiency and subsequently the detection of sequestered sRNAs.

KW - small RNA

KW - sequestration

KW - polyacrylamide gel electrophoresis

KW - denaturation

KW - improved detection

U2 - 10.1007/978-1-4939-6866-4_1

DO - 10.1007/978-1-4939-6866-4_1

M3 - Chapter

SN - 978-1-4939-6864-0

T3 - Methods in Molecular Biology

SP - 1

EP - 6

BT - Microrna detection and target identification

A2 - Dalmay, T

PB - HUMANA PRESS INC

ER -

Improved denaturation of small RNA duplexes and it's application for northern blotting

Abstract / Description of output

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Keywords / Materials (for Non-textual outputs)

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