Comparison of Different Methods for Extraction of Genomic DNA from Ephedra intermedia
Wang Yan
Abstract
Wang Yan
Abstract
Objective:By comparing different extraction methods of genomic DNA,the best extraction method of Ephedra intermedia genomic DNA was selected.Method: Improved cetyltrimethyl ammonium bromide(CTAB) method,SDS method,kit method were used for the extraction of genomic DNA from E.intermedia.The yield and purity of total genomic DNA were examined by agarose gel electrophoresis,UV-spectrophotometry and ISSR-PCR.Result: Three extraction methods could extracted the high amount genomic DNA from fresh tissue of E.intermedia.Improved CTAB method could yield highest purity total DNA,and the electrophoretic bands of amplified products were obvious.On the other hand,SDS method and kit method indicated low purity of the yielded DNA and could not be used in downstream application directly.Conclusion: The improved CTAB method is the best of the three compared methods for extracting the genomic DNA from E.intermedia,which is suitable for PCR reaction and other molecular research in E.intermedia genomic DNA.
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Objective:By comparing different extraction methods of genomic DNA,the best extraction method of Ephedra intermedia genomic DNA was selected.Method: Improved cetyltrimethyl ammonium bromide(CTAB) method,SDS method,kit method were used for the extraction of genomic DNA from E.intermedia.The yield and purity of total genomic DNA were examined by agarose gel electrophoresis,UV-spectrophotometry and ISSR-PCR.Result: Three extraction methods could extracted the high amount genomic DNA from fresh tissue of E.intermedia.Improved CTAB method could yield highest purity total DNA,and the electrophoretic bands of amplified products were obvious.On the other hand,SDS method and kit method indicated low purity of the yielded DNA and could not be used in downstream application directly.Conclusion: The improved CTAB method is the best of the three compared methods for extracting the genomic DNA from E.intermedia,which is suitable for PCR reaction and other molecular research in E.intermedia genomic DNA.
Key concepts: genomic DNA, Agarose gel electrophoresis, DNA extraction, DNA, Chromatography, Extraction (chemistry), Agarose, Molecular biology