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ssr primer set and method for identifying pepper varieties using the primer set

A primer set, pepper technology, applied in biochemical equipment and methods, microbial determination/inspection, recombinant DNA technology, etc., can solve the problems of manpower, material and financial resources, long cycle, etc. Reliable results

Inactive Publication Date: 2015-08-19
贵州省辣椒研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional seed quality inspection methods include field trait identification and grain morphology identification, etc. However, field trait identification requires a long period and consumes a lot of manpower, material and financial resources

Method used

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  • ssr primer set and method for identifying pepper varieties using the primer set
  • ssr primer set and method for identifying pepper varieties using the primer set
  • ssr primer set and method for identifying pepper varieties using the primer set

Examples

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Effect test

Embodiment 1

[0059] (1) Capsicum genomic DNA extraction: select 10 self-cultivation and inbred lines of our research, including H224, H137, H089, H204, H042023A, H009, H015, H004, H102, and H042025D, and collect fresh young leaves of the plants at the seedling stage 2 ~ 3g, using the improved CTAB method to extract DNA samples. The specific steps are: ① Use a 1.5ml centrifuge tube to cover the leaves of the seedlings (about 12-15 mg), and place them on ice; ② Add 100 μl of the extract, and grind them into a slurry; ③ Water bath at 65 °C for 20-40 minutes, during which the centrifuge tubes are inverted twice ; ④Take out the microcentrifuge tube, place it for 5-10min, and cool to room temperature; ⑤Add 100μl of phenol:chloroform:isoamyl alcohol (25:24:1), mix upside down for 5min, and centrifuge at 3200-12000rpm for 10min; ⑥Transfer the supernatant To another 1.5ml centrifuge tube, add 7.5μl 5mol / L NaCl, precipitate DNA with 100μl absolute ethanol (frozen or at room temperature); Dry until ...

Embodiment 2

[0068] (1) Pepper genome DNA extraction: Select 4 pepper varieties: Xiangla No.4, Yuanla Huashuai, Yuanla Hongfeng and Mandi Jintiao, collect 2-3g of fresh young leaves of the plants at the seedling stage, and use the improved CTAB method DNA samples are extracted. The specific steps are: ① Use a 1.5ml centrifuge tube to cover the leaves of the seedlings (about 12-15 mg), and place them on ice; ② Add 100 μl of the extract, and grind them into a slurry; ③ 65 °C water bath for 20-40 minutes, during which the centrifuge tubes are inverted twice ; ④Take out the microcentrifuge tube, place it for 5-10min, and cool to room temperature; ⑤Add 100μl of phenol:chloroform:isoamyl alcohol (25:24:1), mix upside down for 5min, and centrifuge at 3200-12000rpm for 10min; ⑥Transfer the supernatant To another 1.5ml centrifuge tube, add 7.5μl 5mol / L NaCl, precipitate DNA with 100μl absolute ethanol (frozen or at room temperature); Dry until there is no smell of ethanol; ⑧ Add 200 μl sterilized ...

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Abstract

The invention relates to the molecular biological technology field, and concretely relates to an SSR primer group and a method for identification of hot pepper species thereby. The SSR primer group is composed of 20 primers. The method for identification comprises steps: first, DNA extraction of hot pepper leaves; second, PCR amplification; third, electrophoresis detection; fourth, data recording; and fifth, result determination. The primers have high specificity and high amplification efficiency, and can identify different species of hot peppers rapidly. The method is advantaged by short period, low cost, reliable results, manpower saving, material resource saving and land resource saving.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a primer and a method for identifying pepper varieties using the primer. Background technique [0002] Variety identification is an important method in agricultural production, crop breeding and seed inspection, and it is also an important means to protect crop varieties and prevent counterfeit and inferior varieties from entering the market. Traditional seed quality inspection methods include field trait identification and grain morphology identification, etc. However, field trait identification requires a long period and consumes a lot of manpower, material and financial resources. Pepper seed quality monitoring is an important part of pepper production. In the production of peppers, the phenomenon of local species mixing is serious, the routine identification takes a long time, and the field identification is difficult. With the rapid development of molecular biolog...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 何建文苏丹姜虹韩世玉刘永翔刘作易
Owner 贵州省辣椒研究所
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