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Specific probes for identification of asparagus 981 fine variety and its application

A specific and asparagus technology, applied in the field of specific probes for identifying asparagus 981 varieties, can solve the problems of cumbersome operation and long test period

Active Publication Date: 2013-11-20
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Differentiate by determining and measuring growth traits and physiological and biochemical indicators, the test cycle is long and the operation is cumbersome

Method used

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  • Specific probes for identification of asparagus 981 fine variety and its application
  • Specific probes for identification of asparagus 981 fine variety and its application
  • Specific probes for identification of asparagus 981 fine variety and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The specific probe for identifying the improved variety of Asparagus 981 provided by the present invention has the nucleotide sequence described in SEQ ID NO:1 or SEQ ID NO:2.

Embodiment 2

[0042] Asparagus Genomic DNA Extraction

[0043] Extract by CTAB method, weigh 2.5g fresh asparagus, add 5ml CTAB buffer solution and 2g PVPP solid, add liquid nitrogen to grind into powder, and quickly transfer to 10ml plastic centrifuge tube. Incubate at 65°C, shake gently every 5min for several times, centrifuge at 12000r / min for 15min after 30min, carefully absorb the supernatant, add proteinase K to a final concentration of 100μg / ml, 50°C, 3h, mix by inverting from time to time. Extract with equal volume of phenol / chloroform for 10min, centrifuge at 12000rpm for 5min, carefully absorb the supernatant, extract with chloroform for 5min, centrifuge at 12000rpm for 5min, take the supernatant, add 2 / 3 volume of isopropanol, mix it upside down, and store it at -20 Precipitate at ℃ for 15 min, centrifuge at room temperature 12000 rpm for 15 min, remove the supernatant, wash the precipitate with 1 ml of 70% ethanol, centrifuge at 12000 rpm for 15 min, remove the supernatant, and ...

Embodiment 3

[0117] Probes were designed using the two specific fragments obtained in Example 2, and the probe sequences were: SEQ ID NO: 1 and SEQ ID NO: 2. Label the probe and detect the labeling rate according to the instructions of the digoxin labeling kit, prepare the labeled probe and the control according to the operation instructions of the kit, spot each 1 μl of tubes 2-9 on the membrane, let the membrane dry naturally, and bake at 80°C for 2 hours. Add 5ml of buffer I to the membrane in a small beaker [i.e. washing buffer: 0.1M maleic acid, 0.15M NaCl; pH7.5 (20°C); 0.3% (v / v) Tween-20], wash for 5min, pour off Buffer I, add 10ml buffer II [blocking solution, use maleic acid buffer (0.1M maleic acid, 0.15M NaCl, pH7.5) to mix 10×Blocking Solution (tube 6) in a ratio of 1:10 Dilute to 1×working solution], place at room temperature for 30min, pour off buffer II, put the membrane in a petri dish, add 10ml of buffer II to the petri dish, keep at room temperature for 30min, take out t...

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PUM

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Abstract

The invention relates to specific probes for identification of an asparagus 981 fine variety. The specific probe sequences are nucleotide sequences shown as SEQ ID NO:1 or SEQ ID NO:2. The two specific probes obtained in the invention can be used for rapid and accurate identification of the asparagus 981 fine variety.

Description

technical field [0001] The invention relates to the identification of excellent strains of marine algae, in particular to a specific probe for identifying 981 varieties of asparagus and its application. Background technique [0002] The red algae Asparagus is an important agar-producing seaweed. At present, the improved species of Asparagus 981 selected from the wild population in Zhanshan Bay, Qingdao has fast growth, many branches, high temperature resistance, fast growth and stress resistance, high agar content and Well, it has been cultivated for many years in the southern sea area of ​​Fujian, Nan'ao Island, Shantou City, Guangdong Province, etc., and the asparagus cultivation industry has been established in Guangdong, Fujian, Zhejiang, Shandong and Liaoning coastal areas, but it is different from wild asparagus in appearance Difficult to distinguish, it was previously distinguished by measuring the growth traits and physiological and biochemical indicators of the two....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 隋正红王津果周伟常连鹏张淑
Owner OCEAN UNIV OF CHINA
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