Method for extracting genomic DNA from mature and old tung oil tree leaves

A technology of mature leaves and extraction methods, applied in the field of genomic DNA extraction of mature leaves and old leaves of tung tree

Inactive Publication Date: 2012-04-25
WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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  • Abstract
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[0004] After searching, so far, there is no report on the method for extracti

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  • Method for extracting genomic DNA from mature and old tung oil tree leaves
  • Method for extracting genomic DNA from mature and old tung oil tree leaves
  • Method for extracting genomic DNA from mature and old tung oil tree leaves

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Embodiment Construction

[0045] Below in conjunction with accompanying drawing and embodiment describe in detail:

[0046] 1. Extraction method

[0047] Select 10 genotypes of tung tree 10 genotypes of mature leaf mixed fresh samples and old leaf mixed fresh samples, 4 copies each, of which 2 samples were stored for 0 days and 2 samples were stored for 7 months. Use the inventive method to carry out following 8 extraction operations:

[0048] ① Fully grind the leaf tissue sample into powder in liquid nitrogen, quickly weigh 0.3g, quickly transfer it to a 2mL centrifuge tube, and put it in a -70°C refrigerator for use; Stand still for 15min, centrifuge at 12000rpm and discard the supernatant;

[0049] ② Add 500 μl of preheated 3% CTAB extract, mix well, put it in a water bath at 65°C for 40 minutes, centrifuge at 12,000 rpm for 15 minutes at room temperature, and transfer the supernatant to a new 1.5mL centrifuge tube;

[0050] ③The transferred supernatant was extracted once with 24:1 chloroform-iso...

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Abstract

The invention discloses a method for extracting genomic DNA from mature and old tung oil tree leaves, relating to the separation and extraction technology of genomic DNA from plants in the molecular biology experiments. The method mainly comprises the steps of leaf tissue sample pretreatment, cell disruption, non-DNA component extraction, DNA precipitation, ion removal and tissue sample cyclic extraction. The method has the following advantages and positive effects: polyphenols, polysaccharides and other secondary metabolites in which the leaves are rich can be effectively removed by using washing liquid to pretreat the tissue samples of the mature and old tung oil tree leaves once; the step of tissue sample cyclic extraction is absent in the traditional methods for extracting genomic DNA; one tissue sample can undergo genomic DNA extraction four times, thus obviously increasing the yield of the genomic DNA; and the genomic DNA with high quality and high yield can be extracted from the mature and old leaves stored at low temperature for a long time by the method. The method is suitable for extraction of genomic DNA from mature and old tung oil tree leaves.

Description

technical field [0001] The invention relates to a technique for separating and extracting plant genome DNA in molecular biology experiments, in particular to a method for extracting genome DNA from mature leaves and old leaves of tung tree. Background technique [0002] Genomic DNA extraction is an important part of molecular biology experiments. The quality and yield of genomic DNA directly affect the conduct of subsequent DNA-related experiments in molecular biology experiments. Medicinal plants, aromatic plants and woody plants are rich in secondary metabolites; for such plants rich in secondary metabolites, traditional DNA extraction methods are not easy to provide high-quality genomic DNA. The types of secondary metabolites in the leaves of such plants are extremely complex, and the degree of heterogeneity of secondary metabolites among leaves of different leaf ages in different species or even in different seasons of the same species is relatively high; therefore, the...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 彭俊华张玲玲
Owner WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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