Data/hora: 04/06/2024 03:48:36

Provedor de dados: | 69
País: | Chile
Título: | Isolation of high quality RNA from Polyporus umbellatus (Pers.) Fries
Autores: | Zhang,Yuejin; Qin,Yali; Guo,Lijun; Zhou,Zili; Liang,Zongsuo; Zhang,Chenlu; Guo,Hongbo.
Data: | 2012-09-01
Ano: | 2012
Palavras-chave: | Chuling; Medicinal fungus; Polyporus umbellatus; RNA isolation.
Resumo: | 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 &amp;#956;l-1. The Trizol method also showed good quality with the yield 469.60 ng &amp;#956;l-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.
Tipo: | Journal article
Idioma: | Inglês
Identificador: | http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000500010
Editor: | Pontificia Universidad Católica de Valparaíso
Formato: | text/html
Fonte: | Electronic Journal of Biotechnology  v.15 n.5 2012