High-quality DNA extraction method of idesia polycarpa
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An extraction method and high-quality technology, applied in the field of DNA extraction, can solve the problems of small improvement, low DNA concentration, and slow progress in molecular biology research related to S. The effect of fragment distribution is reasonable
Inactive Publication Date: 2019-06-14
RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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However, although the purity of Jacarnia DNA extracted by the kit method has been improved, the improvement is small, and the DNA concentration of each Jacarnia sample is still low. improve
Restricted by this, the research on the related molecular biology of Jatropha is progressing slowly
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Embodiment 1
[0042] Implementation location: Fuyang District, Hangzhou, Zhejiang Province
[0043] Implementation time: April 2018
[0044] At the beginning of April 2018, the tender leaves of three Jacarna japonica samples No. 70, 71, and 72 were collected as materials, and extracted by SDS method (method 1), kit method (method 3), CTAB method (method 3), and the present invention Method (Method 4) Extract the DNA of 3 samples of Jacaranda japonica respectively.
[0045] Wherein, the used chemical reagent of SDS extraction method (method one) comprises:
[0047] 1) Quick-freeze 1 g of plant material in liquid nitrogen, then grind it in a mortar, and transfer the powder to a 2 ml centrifuge tube.
[0048] 2) Add 1.2ml of extraction buffer (3V) preheated to 65°C, shake gently and mix well, add 120μl 20% SDS (to reach a final concentration of 2%), mix we...
Embodiment 2
[0093] Implementation location: Fuyang District, Hangzhou, Zhejiang Province
[0094] Implementation time: April 2018
[0095] At the beginning of April 2018, the young leaves of 6 samples of Jacarina japonica No. 70, 71, 72, 73, 73, and 74 were collected as materials, and the DNA of 5 samples of Jacarnia jacaranda were extracted by the extraction method of the present invention.
[0096] The same place as in Example 1 will not be described in detail, the difference is that the DNA of 6 samples of Jacarna japonica samples are respectively extracted by the extraction method of the present invention as materials, and the SSR typing test and the simplified genome sequencing experiment are carried out respectively, and the results of each test are used to verify the present invention. Whether the Jatropha DNA obtained by the invented extraction method satisfies the requirements of the molecular biology of Jatropha.
[0097] In the SSR typing test, the SSR primer was SSR primer No...
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Abstract
The invention belongs to the technical field of DNA extraction and discloses a high-quality DNA extraction method of idesia polycarpa. According to the method, a CTAB method is taken as a framework, an anti-oxidant is added during grinding, PEG elution is carried out before cracking, subsequent CTAB method operation steps are optimized, so that the high-quality idesia polycarpa DNA extraction method comprising the steps of material collection, material grinding, PEG elution, CTAB cracking, extraction, DNA precipitation, collection and the like is formed. The idesia polycarpa DNA extracted by the method is high in concentration and purity, the quality of the DNA is obviously better than that of an existing method, the average concentration is 438.93 ng / ul, the average concentration is 8.5 times that of a kit method, the OD260 / 280 ratio is 1.94-1.97, the OD260 / 230 ratio is 2.1-2.17, protein contamination and DNA carbohydrate and salt contamination are all well controlled, and the DNA issintact. The examples show that the idesia polycarpa DNA extracted by the method can meet the requirements of molecular tests such as SSR marker typing and genome sequencing simplification; meanwhile,phenol is not used for re-extraction in the method, so that the test steps are simplified, the test time is shortened, and the harm of organic solvents to human health is reduced.
Description
technical field [0001] The invention belongs to the technical field of DNA extraction methods, and in particular relates to a high-quality DNA extraction method of jatropha. Background technique [0002] Jatropha, also known as "oil grape", has strong adaptability, tolerance to drought and barrenness, high fruit yield per plant, whole fruit contains oil, high-quality oil, and has multiple health functions. It is an important woody oil species. Known as the oil tank on the tree. At the same time, the tree has a beautiful shape, the fruit is dark red in autumn, and the cone is drooping. It is also an excellent horticultural ornamental tree species. In recent years, the jatropha industry is showing great potential to start a prairie fire in our country, and it is developing rapidly all over the country. However, the biological characteristics of long childhood, dioecious plants, and many male plants are not conducive to the rapid development of the jatropha industry. It is ur...
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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 徐阳龚榜初吴开云孙维敏江锡兵章平生
Owner RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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