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



Document title: Isolation of high quality RNA from Polyporus umbellatus (Pers.) Fries
Journal: Electronic journal of biotechnology
Database: PERIÓDICA
System number: 000358917
ISSN: 0717-3458
Authors: 1
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Institutions: 1Northwest A & F University, College of Life Sciences, Yangling, Shaanxi. China
Year:
Volumen: 15
Number: 5
Country: Chile
Language: Inglés
Document type: Nota breve o noticia
Approach: Experimental, analítico
English abstract 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
Disciplines: Biología
Keyword: Genética,
Hongos,
Polyporus umbellatus,
Especies medicinales,
ARN,
Biotecnología
Keyword: Biology,
Fungi,
Genetics,
Polyporus umbellatus,
Medicinal species,
RNA,
Biotechnology
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