Comparative analysis of three different methods of total DNA extraction used in Taxus
Gao Lian
Abstract
Gao Lian
Abstract
Species of Taxus in China are endangered and were listed as the national first-class protected wild plant.In present study,three different methods of total DNA extraction,i.e.SDS,CTAB and DNA extraction Kit(DNeasy Plant Mini Kit),were used to isolate genomic DNA from dried leaf materials of Taxus.The total genomic DNA yielded by the three methods were quantified and analyzed by spectrophotometer,agarose gel electrophoresis and PCR reaction respectively.The results of the three methods of DNA extraction were compared and analyzed,which indicated that the CTAB method could yielded relatively pure and high amount total DNA and was highly suited for use directly in downstream applications.The method of DNA extraction using Kit(DNeasy Plant Mini Kit) also could yield high quality total DNA with the highest purity of the DNA comparing to other methods used in this study.However,the relatively lower DNA yield and higher costs of this method might be limited to its wide use.Whereas the SDS method could not yield high quantity of total DNA in most sampled accessions in the present study,which indicated low purity of the yielded DNA of SDS method could not be used in downstream application directly.We also discussed the factors influencing on the total genomic DNA yield and purity,which could help to improve and select the suitable protocol of DNA isolation of plants.
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Species of Taxus in China are endangered and were listed as the national first-class protected wild plant.In present study,three different methods of total DNA extraction,i.e.SDS,CTAB and DNA extraction Kit(DNeasy Plant Mini Kit),were used to isolate genomic DNA from dried leaf materials of Taxus.The total genomic DNA yielded by the three methods were quantified and analyzed by spectrophotometer,agarose gel electrophoresis and PCR reaction respectively.The results of the three methods of DNA extraction were compared and analyzed,which indicated that the CTAB method could yielded relatively pure and high amount total DNA and was highly suited for use directly in downstream applications.The method of DNA extraction using Kit(DNeasy Plant Mini Kit) also could yield high quality total DNA with the highest purity of the DNA comparing to other methods used in this study.However,the relatively lower DNA yield and higher costs of this method might be limited to its wide use.Whereas the SDS method could not yield high quantity of total DNA in most sampled accessions in the present study,which indicated low purity of the yielded DNA of SDS method could not be used in downstream application directly.We also discussed the factors influencing on the total genomic DNA yield and purity,which could help to improve and select the suitable protocol of DNA isolation of plants.
Key concepts: DNA extraction, genomic DNA, Agarose gel electrophoresis, DNA, Chromatography, Extraction (chemistry), Yield (engineering), Taxus