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 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.

Bibliographic Details

Main Authors:	Zhang,Yuejin, Qin,Yali, Guo,Lijun, Zhou,Zili, Liang,Zongsuo, Zhang,Chenlu, Guo,Hongbo
Format:	Digital revista
Language:	English
Published:	Pontificia Universidad Católica de Valparaíso 2012
Online Access:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000500010
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