Abstract
RNA extraction is the foundation of modern molecular biology. It is difficult to extract high quality RNA from grapevine tissues, especially in cases where polysaccharides and polyphenols are rich. In this paper, the modified cetyltrie thylammnonium bromide (CTAB), sodium dodecyl sulphate (SDS) and guanidine isothiocyanate methods were applied to extract high purity and integrity RNA from grapevine green and mature berry skins. Highly-grade RNA was isolated by using modified CTAB method and it is suitable for the demands of further molecular biological research such as reverse-transcription polymerase chain reaction (PCR) (RT-PCR), cDNA library construction and Northern hybridization. Key words: Grapevine berry skins, RNA extraction, reverse-transcription polymerase chain reaction (RT-PCR).
Highlights
Grapevine is one of the most important economic species in the world
The RNA extracted by cetyltrie thylammnonium bromide (CTAB) method has two major ribosomal bands and 28:18 S brightness close to 2:1
The CTAB method set up for RNA extraction from skins of green and mature grapevine berries was compared with two other RNA extraction methods that have been certified to be successfully
Summary
Grapevine is one of the most important economic species in the world. The research of grapevine molecular biology is more and more valuable. Grapevine research requires studies of gene expression and function, which are mainly dependent on RNA quality. A rapid, inexpensive and reliable protocol for the extraction of RNA from grapevine berry skins is challenging because of high concentrations of polysaccharides, polyphenols and other secondary metabolites. Polyphenolics are oxidized quinone compounds which with irreversible binding to RNA (Graham, 1993), results in RNA inactivity and loss in the later extraction with phenol and chloroform (Schneiderbauer et al, 1991; Lin et al, 2003). In the process of extraction to form a common jelly precipitation
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