Method for extracting genomic DNA from young tung oil tree leaves

An extraction method and genome technology, which are applied in the field of genomic DNA extraction from young leaves of tung oil, can solve the problem that there is no report on the extraction method of genomic DNA from young leaves of tung oil

Inactive Publication Date: 2012-04-25
WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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  • Abstract
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Problems solved by technology

[0004] After searching, so far, there is no report on the method for extracting genomic DNA from young leaves of tung tree in the prior art

Method used

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  • Method for extracting genomic DNA from young tung oil tree leaves
  • Method for extracting genomic DNA from young tung oil tree leaves
  • Method for extracting genomic DNA from young 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 2 fresh samples and 3 dry samples of young tung tree leaves mixed samples collected at a growth temperature of around 20°C (November), and 2 mixed samples of young tung tree leaves stored at a temperature of 37°C for 1 year With each two parts of fresh samples of 6 genotype young leaves of tung tree without low temperature storage, use the inventive method to carry out following 19 extraction operations:

[0048] ① Fully grind the leaf tissue samples into powder in liquid nitrogen, quickly weigh 0.3g of fresh leaves, or 0.1g of dry samples, and quickly transfer them to a 2mL centrifuge tube. Put it in a -70°C refrigerator for later use;

[0049] ② Add 1.5mL washing solution, mix well, let stand in ice for 15min, and centrifuge at 12000rpm to discard the supernatant;

[0050] ③After repeating step ② twice, add 500 μl of preheated 3% CTAB extr...

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Abstract

The invention discloses a method for extracting genomic DNA from young 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: proteins, 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 young tung oil tree leaves three times; 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 yield and high quality can be extracted from theyoung leaves stored at low temperature for a long time and from fresh young leaves acquired at different growth temperatures by the method. The method is suitable for extraction of genomic DNA from young tung oil tree leaves.

Description

technical field [0001] The invention relates to a technology for separating and extracting plant genome DNA in molecular biology experiments, in particular to a method for extracting genome DNA from young tung tree leaves. 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 Genomic DNA extr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 彭俊华张玲玲
Owner WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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