Manufacturing method of broccoli variety DNA molecular identity card

A technology of DNA molecules and production methods, applied in the field of molecular biology, can solve problems such as high seed prices, difficulties in variety management and property rights protection, damage to the interests of the country and farmers, and achieve the effect of reducing workload

Pending Publication Date: 2022-03-18
FOOD CROPS RES INST YUNNAN ACADEMY OF AGRI SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, more than 90% of the broccoli seeds used in production are still imported, and the seed prices are extremely high. In the current seed market, phenomena such as "same species with different names", "same name with different species", and "count...

Method used

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  • Manufacturing method of broccoli variety DNA molecular identity card
  • Manufacturing method of broccoli variety DNA molecular identity card
  • Manufacturing method of broccoli variety DNA molecular identity card

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0035] Example 1 Production method of "axi green" DNA molecular ID card

[0036] (1) Blue and chlorist genome DNA extraction:

[0037] Take the same green vane 200-300 mg placed in 2.0 ml of centrifuge tube, and the liquid is fully grinded. 700 μL of a CTAB extract of 65 ° C for 65 ° C was added, mixed with a mixture of 65 ° C for 60 min and 2-3 times. Emorbent dichloromethane and isotenthane mixed liquid per tube were added, and dichloromethane in the mixture was 24: 1, and the volume ratio of isopentyl alcohol was 24: 1, which was allowed to stand for 10 min after mixing and centrifuged at 12000 rpm for 15 min. The supernatant was transferred to a new centrifuge tube, adding an equal volume of 0. ° C pre-cooled isopropanol, reversing mix, 30 min after 30 min at 4 ° C, 12000 rpm for 10min, discard the supernatant, add 70% V / V ethanol, rotated 2-3 times, abandon the ethanol solution, and turned onto the test of the pad with filter paper, and placed over 10 min. 100 μl of ultrapu...

Embodiment 2

[0062] Example 2 Made of DNA molecular ID cards of the remaining 21 blue calendar plants

[0063] Example 2 Except as in Example 1 except that the varieties used, the remaining experimental operation steps were the same as in Example 1, and will not be described again. The 22 green-chop variety molecular fingerprint information is shown in Table 2. According to the DNA molecular fingerprint information of the blue and white owners, the corresponding basic product information data is reselected, and the DNA molecular ID card can be produced according to the method described in Example 1.

[0064] Table 2 22 green-choppy variety molecular fingerprint information

[0065]

[0066]

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PUM

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Abstract

The invention relates to a method for manufacturing a broccoli variety DNA (Deoxyribonucleic Acid) molecular identity card, which comprises the following steps of: screening four pairs of core primers, performing genotyping on 22 current large-area mainly-cultivated broccoli varieties, manufacturing a two-dimensional code broccoli variety DNA molecular identity card, and identifying the broccoli varieties by using fewest primers with wide genome coverage. And the broccoli varieties which are mainly cultivated at present can be distinguished one by one, so that the complicated workload and the high detection cost are reduced.

Description

Technical field [0001] The present invention belongs to the field of molecular biology, and is specifically a method based on the authenticity identification of blue-chop-based varieties developed by SSR marking technology, and is used in the identification and variety management of variety molecules. Background technique [0002] Brassica Olecea L.Var.italic, also known as a variety of cabbage vegetables originated in the European Mediterranean Coast. Blue-choppy is favored by consumers at home and abroad due to anti-cancer active ingredients. The demand and planting area of ​​blue-choppy is also increased year by year. However, in my country's 30 years of green and chuck cultivation history, its production species mainly depends on foreign imports. At present, more than 90% of the green-choppy vegetables are still imported, and the seed is very high. The current seed market "the same kind of" same kind "," same name "," counterfeit package ", etc. , There is a vacation and sale...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/6895C12Q1/686C12Q2600/156C12Q2565/125C12Q2563/107
Inventor 管俊娇张建华丁燕木万福张鹏李社萍杨龙杨晓洪黄清梅李彦刚
Owner FOOD CROPS RES INST YUNNAN ACADEMY OF AGRI SCI
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