METHOD FOR EXTRACTING HIGH-QUALITY DNA FROM IDESIA POLYCARPA
Abstract The present invention belongs to the technical field of DNA extraction, and discloses a method for extracting high-quality DNA from Idesia polycarpa. The present invention adopts a CTAB method as a framework, adds an antioxidant during grinding, carries out PEG elution before lysis, and opt...
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| creator | Wu, Kaiyun Jiang, Xibing Sun, Weimin Xu, Yang Gong, Bangchu Zhang, Pingsheng |
| description | Abstract The present invention belongs to the technical field of DNA extraction, and discloses a method for extracting high-quality DNA from Idesia polycarpa. The present invention adopts a CTAB method as a framework, adds an antioxidant during grinding, carries out PEG elution before lysis, and optimizes the subsequent operation steps of the CTAB method to form a method for extracting high-quality DNA from Idesia polycarpa, which includes the steps of material collection, material grinding, PEG elution, CTAB lysis, extraction, and DNA precipitation and collection. The Idesia polycarpa DNA extracted by the method is high in concentration and high in purity and has better quality than that of the existing method. An average concentration is 438.93 ng/ul, which is 8.5 times of that obtained by a kit method. An OD260/280 ratio is 1.94-1.97, and an OD260/230 ratio is 21.-2.17. The protein pollution and DNA saccharides and salt pollution are well controlled, and DNA is complete. Embodiments show that the Idesia polycarpa DNA extracted by the method can meet the requirements of molecular test such as SSR marker typing and reduced-representation sequencing. Meanwhile, the method does not use phenol for re-extraction, so that while the test time is shortened, and the test steps are simplified, the harm of organic solvents on the health of human body can also be reduced. Drawings of Description 23130bp+ 9416bp + 6557bp + 4361bp + 2322bp + 2027bp +9 Fig. 1 |
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The present invention adopts a CTAB method as a framework, adds an antioxidant during grinding, carries out PEG elution before lysis, and optimizes the subsequent operation steps of the CTAB method to form a method for extracting high-quality DNA from Idesia polycarpa, which includes the steps of material collection, material grinding, PEG elution, CTAB lysis, extraction, and DNA precipitation and collection. The Idesia polycarpa DNA extracted by the method is high in concentration and high in purity and has better quality than that of the existing method. An average concentration is 438.93 ng/ul, which is 8.5 times of that obtained by a kit method. An OD260/280 ratio is 1.94-1.97, and an OD260/230 ratio is 21.-2.17. The protein pollution and DNA saccharides and salt pollution are well controlled, and DNA is complete. Embodiments show that the Idesia polycarpa DNA extracted by the method can meet the requirements of molecular test such as SSR marker typing and reduced-representation sequencing. Meanwhile, the method does not use phenol for re-extraction, so that while the test time is shortened, and the test steps are simplified, the harm of organic solvents on the health of human body can also be reduced. 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The present invention adopts a CTAB method as a framework, adds an antioxidant during grinding, carries out PEG elution before lysis, and optimizes the subsequent operation steps of the CTAB method to form a method for extracting high-quality DNA from Idesia polycarpa, which includes the steps of material collection, material grinding, PEG elution, CTAB lysis, extraction, and DNA precipitation and collection. The Idesia polycarpa DNA extracted by the method is high in concentration and high in purity and has better quality than that of the existing method. An average concentration is 438.93 ng/ul, which is 8.5 times of that obtained by a kit method. An OD260/280 ratio is 1.94-1.97, and an OD260/230 ratio is 21.-2.17. The protein pollution and DNA saccharides and salt pollution are well controlled, and DNA is complete. Embodiments show that the Idesia polycarpa DNA extracted by the method can meet the requirements of molecular test such as SSR marker typing and reduced-representation sequencing. Meanwhile, the method does not use phenol for re-extraction, so that while the test time is shortened, and the test steps are simplified, the harm of organic solvents on the health of human body can also be reduced. 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The present invention adopts a CTAB method as a framework, adds an antioxidant during grinding, carries out PEG elution before lysis, and optimizes the subsequent operation steps of the CTAB method to form a method for extracting high-quality DNA from Idesia polycarpa, which includes the steps of material collection, material grinding, PEG elution, CTAB lysis, extraction, and DNA precipitation and collection. The Idesia polycarpa DNA extracted by the method is high in concentration and high in purity and has better quality than that of the existing method. An average concentration is 438.93 ng/ul, which is 8.5 times of that obtained by a kit method. An OD260/280 ratio is 1.94-1.97, and an OD260/230 ratio is 21.-2.17. The protein pollution and DNA saccharides and salt pollution are well controlled, and DNA is complete. Embodiments show that the Idesia polycarpa DNA extracted by the method can meet the requirements of molecular test such as SSR marker typing and reduced-representation sequencing. Meanwhile, the method does not use phenol for re-extraction, so that while the test time is shortened, and the test steps are simplified, the harm of organic solvents on the health of human body can also be reduced. Drawings of Description 23130bp+ 9416bp + 6557bp + 4361bp + 2322bp + 2027bp +9 Fig. 1</abstract><oa>free_for_read</oa></addata></record> |
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| subjects | BEER BIOCHEMISTRY CHEMISTRY COMPOSITIONS THEREOF CULTURE MEDIA ENZYMOLOGY METALLURGY MICROBIOLOGY MICROORGANISMS OR ENZYMES MUTATION OR GENETIC ENGINEERING PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS SPIRITS VINEGAR WINE |
| title | METHOD FOR EXTRACTING HIGH-QUALITY DNA FROM IDESIA POLYCARPA |
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