Extraction of genomic DNA from polysaccharide- and phenolics-rich Ginkgo biloba

Abstract

One prerequisite to reliable molecular biology work is that the genomic DNA of a sample should be of good quality. The isolation of intact, high-molecular-mass genomic DNA is essential for many molecular biology applications including long Polymerase Chain Reaction (PCR), endonuclease restriction digestion, Southern blot analysis, and genomic library construction. Many protocols are available for the extraction of DNA from plant material. DNA extraction of Ginkgo biloba is quite difficult to work on because of the high phenolic and polysaccharide content of its leaves. This study aimed to determine which protocol to use and which part of Ginkgo tree is most appropriate to extract good-quality genomic DNA. For this purpose, cetyltrimethylammonium bromide protocol and protocol of commercially available kit by EZ1 Nucleic acid isolation system have been optimized for extraction of genomic DNA from G. biloba leaves. Efficient yields of high-quality amplifiable DNA was produced rapidly with kit by EZ1 Nucleic acid isolation method. The purified DNA which has excellent spectral quality was efficiently amplified by 5 arbitrary primers (OPA11-15), and was suitable for long-fragment PCR amplification. Key words: Genomic DNA, random amplification of polymorphic DNA polymerase chain reaction, DNA extraction, phenolics.