Abstract:

Isolation of high-quality RNA from ginkgo (Ginkgo biloba L.) kernels is a prerequisite to acquire the pharmaceutical protein genes and study the development and maturation of ginkgo seeds at the gene expression level. Ginkgo seeds are rich in protein , polysaccharide and phenolic compounds , which increases the difficulties in isolating RNA from them. In this study, modified CTAB method and Trizol single-step method were combined and the extraction conditions were optimized for RNA extraction. The quality of total RNA was analyzed with agarose gel electrophoresis and UV spectrophotometer. The bright and clear strip of 28S rRNA and 18S rRNA were shown in the RNA electrophoresis, and the luminance signal ratio was approximate 2：1. The value of OD260/OD280 was 1.85 to 2.00. It is indicated that the method could isolate high quality and high integrity total RNA. When the acquired total RNA was used in the molecular biology experiment, sharp target hybridization signals were obtained from Northern blotting, and the target gene fragments were also successfully amplified by RT-PCR and 5'-RACE. These results also demonstrated that the total RNA isolated by this method was of sufficient quality for subsequent molecular applications.