A method for obtaining high quality RNA from paraffin sections of plant tissues by laser microdissection

Hirokazu Takahashi, Hisae Kamakura, Yutaka Sato, Katsuhiro Shiono, Tomomi Abiko, Nobuhiro Tsutsumi, Yoshiaki Nagamura, Naoko K. Nishizawa, Mikio Nakazono

研究成果: ジャーナルへの寄稿記事

70 引用 (Scopus)

抄録

Laser microdissection (LM) combined with microarray analysis or next-generation sequencing of cDNA is a powerful tool for understanding molecular events in individual cell types of plants as well as animals. Obtaining high quality RNA is essential for this approach. For plant tissues, paraffin-embedded sections better preserve cell structure than do frozen sections. However, the conventional method for preparing paraffin sections is a lengthy process involving embedding the tissue and floating and drying the sections, during which time RNA degradation occurs. Here, we describe a method for preparing serial sections that greatly reduces RNA degradation: we reduced (1) the embedding time from 4-6 days to about 5 h by using a recently developed microwave method; (2) the time of floating sections from ~10 min to less than 5 min, (3) the drying time from ~12 to 1 h; and (4) the drying temperature from 42 to 4°C. With this method, we were able to isolate higher integrity RNA from many kinds of plant tissues than is typically obtained by the conventional paraffin preparation method. The improvement in RNA quality and yield removes a major obstacle to the widespread use of LM with high-throughput technologies for plants.

元の言語英語
ページ(範囲)807-813
ページ数7
ジャーナルJournal of Plant Research
123
発行部数6
DOI
出版物ステータス出版済み - 6 1 2010
外部発表Yes

Fingerprint

alkanes
lasers
plant tissues
RNA
drying
methodology
degradation
cell structures
drying temperature
preserves
animals
cells

All Science Journal Classification (ASJC) codes

  • Plant Science

これを引用

A method for obtaining high quality RNA from paraffin sections of plant tissues by laser microdissection. / Takahashi, Hirokazu; Kamakura, Hisae; Sato, Yutaka; Shiono, Katsuhiro; Abiko, Tomomi; Tsutsumi, Nobuhiro; Nagamura, Yoshiaki; Nishizawa, Naoko K.; Nakazono, Mikio.

:: Journal of Plant Research, 巻 123, 番号 6, 01.06.2010, p. 807-813.

研究成果: ジャーナルへの寄稿記事

Takahashi, H, Kamakura, H, Sato, Y, Shiono, K, Abiko, T, Tsutsumi, N, Nagamura, Y, Nishizawa, NK & Nakazono, M 2010, 'A method for obtaining high quality RNA from paraffin sections of plant tissues by laser microdissection', Journal of Plant Research, 巻. 123, 番号 6, pp. 807-813. https://doi.org/10.1007/s10265-010-0319-4
Takahashi, Hirokazu ; Kamakura, Hisae ; Sato, Yutaka ; Shiono, Katsuhiro ; Abiko, Tomomi ; Tsutsumi, Nobuhiro ; Nagamura, Yoshiaki ; Nishizawa, Naoko K. ; Nakazono, Mikio. / A method for obtaining high quality RNA from paraffin sections of plant tissues by laser microdissection. :: Journal of Plant Research. 2010 ; 巻 123, 番号 6. pp. 807-813.
@article{ced3f95d781a4ad5886eccbc1acd60e1,
title = "A method for obtaining high quality RNA from paraffin sections of plant tissues by laser microdissection",
abstract = "Laser microdissection (LM) combined with microarray analysis or next-generation sequencing of cDNA is a powerful tool for understanding molecular events in individual cell types of plants as well as animals. Obtaining high quality RNA is essential for this approach. For plant tissues, paraffin-embedded sections better preserve cell structure than do frozen sections. However, the conventional method for preparing paraffin sections is a lengthy process involving embedding the tissue and floating and drying the sections, during which time RNA degradation occurs. Here, we describe a method for preparing serial sections that greatly reduces RNA degradation: we reduced (1) the embedding time from 4-6 days to about 5 h by using a recently developed microwave method; (2) the time of floating sections from ~10 min to less than 5 min, (3) the drying time from ~12 to 1 h; and (4) the drying temperature from 42 to 4°C. With this method, we were able to isolate higher integrity RNA from many kinds of plant tissues than is typically obtained by the conventional paraffin preparation method. The improvement in RNA quality and yield removes a major obstacle to the widespread use of LM with high-throughput technologies for plants.",
author = "Hirokazu Takahashi and Hisae Kamakura and Yutaka Sato and Katsuhiro Shiono and Tomomi Abiko and Nobuhiro Tsutsumi and Yoshiaki Nagamura and Nishizawa, {Naoko K.} and Mikio Nakazono",
year = "2010",
month = "6",
day = "1",
doi = "10.1007/s10265-010-0319-4",
language = "English",
volume = "123",
pages = "807--813",
journal = "Journal of Plant Research",
issn = "0918-9440",
publisher = "Springer Japan",
number = "6",

}

TY - JOUR

T1 - A method for obtaining high quality RNA from paraffin sections of plant tissues by laser microdissection

AU - Takahashi, Hirokazu

AU - Kamakura, Hisae

AU - Sato, Yutaka

AU - Shiono, Katsuhiro

AU - Abiko, Tomomi

AU - Tsutsumi, Nobuhiro

AU - Nagamura, Yoshiaki

AU - Nishizawa, Naoko K.

AU - Nakazono, Mikio

PY - 2010/6/1

Y1 - 2010/6/1

N2 - Laser microdissection (LM) combined with microarray analysis or next-generation sequencing of cDNA is a powerful tool for understanding molecular events in individual cell types of plants as well as animals. Obtaining high quality RNA is essential for this approach. For plant tissues, paraffin-embedded sections better preserve cell structure than do frozen sections. However, the conventional method for preparing paraffin sections is a lengthy process involving embedding the tissue and floating and drying the sections, during which time RNA degradation occurs. Here, we describe a method for preparing serial sections that greatly reduces RNA degradation: we reduced (1) the embedding time from 4-6 days to about 5 h by using a recently developed microwave method; (2) the time of floating sections from ~10 min to less than 5 min, (3) the drying time from ~12 to 1 h; and (4) the drying temperature from 42 to 4°C. With this method, we were able to isolate higher integrity RNA from many kinds of plant tissues than is typically obtained by the conventional paraffin preparation method. The improvement in RNA quality and yield removes a major obstacle to the widespread use of LM with high-throughput technologies for plants.

AB - Laser microdissection (LM) combined with microarray analysis or next-generation sequencing of cDNA is a powerful tool for understanding molecular events in individual cell types of plants as well as animals. Obtaining high quality RNA is essential for this approach. For plant tissues, paraffin-embedded sections better preserve cell structure than do frozen sections. However, the conventional method for preparing paraffin sections is a lengthy process involving embedding the tissue and floating and drying the sections, during which time RNA degradation occurs. Here, we describe a method for preparing serial sections that greatly reduces RNA degradation: we reduced (1) the embedding time from 4-6 days to about 5 h by using a recently developed microwave method; (2) the time of floating sections from ~10 min to less than 5 min, (3) the drying time from ~12 to 1 h; and (4) the drying temperature from 42 to 4°C. With this method, we were able to isolate higher integrity RNA from many kinds of plant tissues than is typically obtained by the conventional paraffin preparation method. The improvement in RNA quality and yield removes a major obstacle to the widespread use of LM with high-throughput technologies for plants.

UR - http://www.scopus.com/inward/record.url?scp=78049306506&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78049306506&partnerID=8YFLogxK

U2 - 10.1007/s10265-010-0319-4

DO - 10.1007/s10265-010-0319-4

M3 - Article

C2 - 20221666

AN - SCOPUS:78049306506

VL - 123

SP - 807

EP - 813

JO - Journal of Plant Research

JF - Journal of Plant Research

SN - 0918-9440

IS - 6

ER -