An improved and reproducible protocol for the extraction of high quality fungal RNA from plant biomass substrates

被引：14

作者：

Patyshakuliyeva, Aleksandrina ^{[1
,2
]}

Makela, Miia R. ^{[3
]}

Sietio, Outi-Maaria ^{[3
]}

de Vries, Ronald P. ^{[1
,2
]}

Hilden, Kristiina S. ^{[3
]}

机构：

[1] CBS KNAW Fungal Biodivers Ctr, NL-3584 CT Utrecht, Netherlands

[2] Univ Utrecht, Utrecht, Netherlands

[3] Univ Helsinki, Dept Food & Environm Sci, FIN-00014 Helsinki, Finland

来源：

FUNGAL GENETICS AND BIOLOGY | 2014年 / 72卷

关键词：

Lignocellulose (plant biomass); RNA extraction; Agaricus bisporus; Dichomitus squalens; CsCl gradient ultracentrifugation; Gene expression; MESSENGER-RNA; RIBONUCLEIC-ACID; HUMIC SUBSTANCES; LIGNIN; DNA; QUANTIFICATION; FERMENTATION; ENZYMES; STATE;

D O I：

10.1016/j.fgb.2014.06.001

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

Isolation of high quantity and quality RNA is a crucial step in the detection of meaningful gene expression data. Obtaining intact fungal RNA from complex lignocellulosic substrates is often difficult, producing low integrity RNA which perform poorly in downstream applications. In this study we developed an RNA extraction method using CsCl centrifugation procedure, modified from previous reports and adapted for isolation of RNA from plant biomass. This method provided high level of integrity and good quantity of RNA which were suitable for reliable analyses of gene expression and produced consistent and reproducible results. (C) 2014 Elsevier Inc. All rights reserved.

引用

页码：201 / 206

页数：6

共 35 条

[1]

Aiken G. R., HUMIC SUBSTANCES SOI

[2] The presence of humic substances and DNA in RNA extracts affects hybridization results [J].

Alm, EW ;

Zheng, DD ;

Raskin, L .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2000, 66 (10) :4547-4554

[3] Effects of humic substances on fluorometric DNA quantification and DNA hybridization [J].

Bachoon, DS ;

Otero, E ;

Hodson, RE .

JOURNAL OF MICROBIOLOGICAL METHODS, 2001, 47 (01) :73-82

[4]

Chang S. J., 1993, Plant Molecular Biology Reporter, V11, P113, DOI 10.1007/BF02670468

[5] ISOLATION OF BIOLOGICALLY-ACTIVE RIBONUCLEIC-ACID FROM SOURCES ENRICHED IN RIBONUCLEASE [J].

CHIRGWIN, JM ;

PRZYBYLA, AE ;

MACDONALD, RJ ;

RUTTER, WJ .

BIOCHEMISTRY, 1979, 18 (24) :5294-5299

[6] Deconstruction of Lignocellulosic Biomass to Fuels and Chemicals [J].

Chundawat, Shishir P. S. ;

Beckham, Gregg T. ;

Himmel, Michael E. ;

Dale, Bruce E. .

ANNUAL REVIEW OF CHEMICAL AND BIOMOLECULAR ENGINEERING, VOL 2, 2011, 2 :121-145

[7] Fungal Bioconversion of Lignocellulosic Residues; Opportunities & Perspectives [J].

Dashtban, Mehdi ;

Schraft, Heidi ;

Qin, Wensheng .

INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES, 2009, 5 (06) :578-595

[8]

England LS, 2003, RIV BIOL-BIOL FORUM, V96, P317

[9] Extraction and detection of baculoviral DNA from lake water, detritus and forest litter [J].

England, LS ;

Trevors, JT ;

Holmes, SB .

JOURNAL OF APPLIED MICROBIOLOGY, 2001, 90 (04) :630-636

[10]

Fleige Simone, 2006, Molecular Aspects of Medicine, V27, P126, DOI 10.1016/j.mam.2005.12.003

← 1 2 3 4 →