#Northern blot free#

Here we report the application of a CTAB- based method for RNA extraction using an additional clean-up purification step. procedure to improve RNA quality for root gene expression studies.Д¬onventional RNA extraction methods using CTAB are not suitable for gene expression studies and need to be improved. Hence, we have developed a new extraction. However, obtaining high quality RNA from grapevine roots under hydroponic culture is difficult. Identification of candidate genes involved in the tolerance to abiotic stress is becoming a crucial strategy to select and breed resilient genotypes. Grapevine root system plays a great role in sensing and adapting to abiotic and biotic stresses. We provide an efficient and low-cost method for extracting high quantities of high-quality RNA from plants that are rich in polyphenols and polysaccharides, and this method was also validated for the isolation of high-quality RNA from other plants. Furthermore, to confirm downstream amenability, the high-quality RNA obtained from this method was successfully used for RT-PCR, real-time RT-PCR and Northern blot analysis. The method of RNA extraction presented here was superior to the conventional CTAB method in terms of RNA isolation efficiency and was more sample-adaptable and cost-effective than commercial kits.

#Northern blot free#

Spectrophotometric analysis showed that A260/280 ratios of RNA samples were in the range of 2.11 to 2.13, and A260/230 ratios were in the range of 2.04-2.24, indicating that the RNA samples were free of polyphenols, polysaccharides, and protein contaminants after isolation. RNA quality was measured for yield and purity, and the proposed protocol yielded high quantities of RNA (10.1-12.9 Ојg/g high-quality RNA from several different kenaf tissues. In the present study, we proposed a simplified, time-saving and low-cost extraction method for isolating high quantities of. However, high levels of polysaccharide and polyphenol compounds in kenaf tissues could irreversibly bind to and coprecipitate with RNA, which complicates RNA extraction. The isolation of high-quality RNA from kenaf is crucial for genetic and molecular biology studies.