Method for obtaining transgenic alfalfa and special expression vector CPB-LAR-GFP thereof
A CPB-LAR-GFP, transgenic alfalfa technology, applied in the direction of introducing foreign genetic material, genetic engineering, plant genetic improvement using vectors, etc., to avoid the increase of production costs, reduce pollution, and reduce the pressure of environmental pollution.
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[0045] 1. Extraction of total RNA of Ormosia japonica:
[0046] According to the characteristics of red bean grass rich in polyphenols, polysaccharides and secondary metabolites, the fresh young tissue of Gansu red bean grass plant was used to prepare RNA crude extract by CTAB method, and the extracted RNA was further purified to find out a suitable Extraction method of total RNA of Ormosia japonica. This method mainly prevents secondary metabolites from combining with RNA by adding 4% PVP to the extraction buffer; adds 2% β-mercaptoethanol to reduce the interference of polyphenols; utilizes water-saturated phenol: chloroform: isoamyl alcohol to extract Removing the interference of DNA, protein, chlorophyll, anthocyanin and other pigments on RNA not only saves experimental costs, but also reduces the probability of RNA degradation and improves the quality of RNA. RNA is efficiently separated from polysaccharides by selective precipitation of RNA using LiCl. Purification of t...
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