Safer DNA extraction from plant tissues using sucrose buffer and glass fiber filter.
For some plant species, DNA extraction and downstream experiments are inhibited by various chemicals such as polysaccharides and polyphenols. This short communication proposed an organic-solvent free (except for ethanol) extraction method. This method consists of an initial washing step with STE buffer (0.25 M sucrose, 0.03 M Tris, 0.05 M EDTA), followed by DNA extraction using a piece of glass fiber filter. The advantages of this method are its safety and low cost. The purity of the DNA solution obtained using this method is not necessarily as high as that obtained using the STE/CTAB method, but it is sufficient for PCR experiments. These points were demonstrated empirically with two species, Japanese speedwell and common dandelion, for which DNA has proven difficult to amplify via PCR in past studies.
Osaka City Institute of Public Health and Environmental Sciences, Osaka, Japan. takakura@nature.email.ne.jp
Koh-Ichi
K
Takakura
Takayuki
T
Nishio
D002021:Buffers; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D000431:Ethanol / Q000737:chemistry; D005898:Glass / Q000737:chemistry*; D016133:Polymerase Chain Reaction / Q000379:methods; D012997:Solvents / Q000737:chemistry; D013395:Sucrose / Q000737:chemistry*; D031205:Taraxacum / Q000737:chemistry / Q000235:genetics*; D032264:Veronica / Q000737:chemistry / Q000235:genetics*
D002021:Buffers; D018744:DNA, Plant; D012997:Solvents; C009753:fiberglass; D000431:Ethanol; D013395:Sucrose