Isolation of high quality RNA from Polyporus umbellatus (Pers.) Fries

Background: The dried sclerotium of medicinal fungus Polyporus umbellatus (Pers.) Fries has many pharmacological functions such as diuretic and anticancer activity, in which high-content polysaccharides may play an important role. However, RNA isolation is difficult in filamentous fungi and lacking...

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Autores principales:	Zhang,Yuejin, Qin,Yali, Guo,Lijun, Zhou,Zili, Liang,Zongsuo, Zhang,Chenlu, Guo,Hongbo
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2012
Materias:	Chuling medicinal fungus Polyporus umbellatus RNA isolation
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000500010
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spelling	oai:scielo:S0717-345820120005000102012-11-29Isolation of high quality RNA from Polyporus umbellatus (Pers.) FriesZhang,YuejinQin,YaliGuo,LijunZhou,ZiliLiang,ZongsuoZhang,ChenluGuo,Hongbo Chuling medicinal fungus Polyporus umbellatus RNA isolation Background: The dried sclerotium of medicinal fungus Polyporus umbellatus (Pers.) Fries has many pharmacological functions such as diuretic and anticancer activity, in which high-content polysaccharides may play an important role. However, RNA isolation is difficult in filamentous fungi and lacking in P. umbellatus. Results: Five methods for RNA extraction from five strains collected from four provinces were assessed for their ability to recover a high-quality RNA applicable for sequence-related amplification polymorphism (SRAP) PCR and GDP-D-mannose pyrophosphorylase (GMP) gene expression profiles. Both A260/A280 and A260/A230 ratios of the best Trizol Plus + RNAiso-mate for Plant Tissue method are around 2 with a yield of 1122.00 ± 0.21 ng μl-1. The Trizol method also showed good quality with the yield 469.60 ng μl-1. The SRAP PCR amplified clear and polymorphic bands in all five cDNA samples transcribed from RNA by using primer Me4-Em4. GMP gene fragment (1251 bp) was successfully amplified by RT-PCR, suggesting the integrity of isolated RNA. Conclusion: All these results showed that the total RNA isolated by this protocol is of sufficient quality for subsequent molecular applications.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.15 n.5 20122012-09-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000500010en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Chuling medicinal fungus Polyporus umbellatus RNA isolation
spellingShingle	Chuling medicinal fungus Polyporus umbellatus RNA isolation Zhang,Yuejin Qin,Yali Guo,Lijun Zhou,Zili Liang,Zongsuo Zhang,Chenlu Guo,Hongbo Isolation of high quality RNA from Polyporus umbellatus (Pers.) Fries
description	Background: The dried sclerotium of medicinal fungus Polyporus umbellatus (Pers.) Fries has many pharmacological functions such as diuretic and anticancer activity, in which high-content polysaccharides may play an important role. However, RNA isolation is difficult in filamentous fungi and lacking in P. umbellatus. Results: Five methods for RNA extraction from five strains collected from four provinces were assessed for their ability to recover a high-quality RNA applicable for sequence-related amplification polymorphism (SRAP) PCR and GDP-D-mannose pyrophosphorylase (GMP) gene expression profiles. Both A260/A280 and A260/A230 ratios of the best Trizol Plus + RNAiso-mate for Plant Tissue method are around 2 with a yield of 1122.00 ± 0.21 ng μl-1. The Trizol method also showed good quality with the yield 469.60 ng μl-1. The SRAP PCR amplified clear and polymorphic bands in all five cDNA samples transcribed from RNA by using primer Me4-Em4. GMP gene fragment (1251 bp) was successfully amplified by RT-PCR, suggesting the integrity of isolated RNA. Conclusion: All these results showed that the total RNA isolated by this protocol is of sufficient quality for subsequent molecular applications.
author	Zhang,Yuejin Qin,Yali Guo,Lijun Zhou,Zili Liang,Zongsuo Zhang,Chenlu Guo,Hongbo
author_facet	Zhang,Yuejin Qin,Yali Guo,Lijun Zhou,Zili Liang,Zongsuo Zhang,Chenlu Guo,Hongbo
author_sort	Zhang,Yuejin
title	Isolation of high quality RNA from Polyporus umbellatus (Pers.) Fries
title_short	Isolation of high quality RNA from Polyporus umbellatus (Pers.) Fries
title_full	Isolation of high quality RNA from Polyporus umbellatus (Pers.) Fries
title_fullStr	Isolation of high quality RNA from Polyporus umbellatus (Pers.) Fries
title_full_unstemmed	Isolation of high quality RNA from Polyporus umbellatus (Pers.) Fries
title_sort	isolation of high quality rna from polyporus umbellatus (pers.) fries
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2012
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000500010
work_keys_str_mv	AT zhangyuejin isolationofhighqualityrnafrompolyporusumbellatuspersfries AT qinyali isolationofhighqualityrnafrompolyporusumbellatuspersfries AT guolijun isolationofhighqualityrnafrompolyporusumbellatuspersfries AT zhouzili isolationofhighqualityrnafrompolyporusumbellatuspersfries AT liangzongsuo isolationofhighqualityrnafrompolyporusumbellatuspersfries AT zhangchenlu isolationofhighqualityrnafrompolyporusumbellatuspersfries AT guohongbo isolationofhighqualityrnafrompolyporusumbellatuspersfries
_version_	1718441861770641408

Isolation of high quality RNA from Polyporus umbellatus (Pers.) Fries

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