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PCR identification method for sorghum halepense and similar species and kit

A technology of sorghum and black sorghum, which is applied in the field of interspecific identification of quarantine weeds Pseudosorghum and its similar species, which can solve the problems of narrow identification range and long time consumption

Inactive Publication Date: 2010-06-23
INSPECTION & QUARANTINE TECH CENT OF FUJIAN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The identification range of this identification method is limited to two species of pseudosorghum and pseudosorghum, narrower than the identification range of the present invention, and takes about 1 times longer

Method used

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  • PCR identification method for sorghum halepense and similar species and kit
  • PCR identification method for sorghum halepense and similar species and kit
  • PCR identification method for sorghum halepense and similar species and kit

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Embodiment 1

[0067] 1 Materials and methods

[0068] 1.1 Experimental materials

[0069] False sorghum, black sorghum, silk sorghum, pseudosorghum, and Sudan grass seeds are provided by Fujian Entry-Exit Inspection and Quarantine Bureau.

[0070] 1.2 DNA extraction

[0071] Take 0.05g of sample, freeze it in liquid nitrogen and grind it to powder, add 500μl TES * , add 50-100 μg proteinase K, incubate at 55-60 °C for 0.5-1 h, shake gently during the mixing period; adjust the NaCl concentration to 1.4 mol / L, add 1 / 10 volume of 10% CTAB, 65 °C for 10 min; add 1 volume Chloroform:isoamyl alcohol (24:1), shake and mix well, at 0°C for 30min, then centrifuge at 12000rpm for 10min at 4°C; take the supernatant, add 225μl 5mol NH 4 AC, mix well, put on ice for more than 30min, centrifuge at 12000rpm at 4°C for 10min; take the supernatant, add 3 μl RNase, centrifuge at 37°C for 15min, 4°C 12000 for 10min; take the supernatant, add 0.55 volume of isopropanol to precipitate DNA, and immediately C...

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Abstract

The invention relates to identification filed of importing quarantine weed species level, in particular to an identification method for sorghum halepense, black sorghum, and similar specicies and a kit, silk sorghum, sorghum propinquum, and sorghum sudanense these five species and the kit used by the method. The identification method comprises: 1, identifying PCR; 2, electrophoresis detection. The kit comprises 3 pairs of PCR reaction primers and a standard map. The advantages of the invention are as follows: the invention can solves the problem of identification of the sorghum halepense and the similar species of the sorghum halepense, and a kit which completes molecular identification of the sorghum halepense and the similar species of the sorghum halepense in 8 hours is provided.

Description

technical field [0001] The invention relates to the technical field of interspecific identification of quarantine weeds pseudosorghum and its similar species, and relates to a PCR method for rapid identification of pseudosorghum, black sorghum, silk sorghum, pseudosorghum and Sudan grass and the method used in the method. the kit. Background technique [0002] False sorghum is one of the top ten malignant weeds in the world, and my country has included it in the list of harmful organisms. The characteristics of the seeds of pseudosorghum are very similar to those of other species of the genus Sorghum, such as pseudosorghum and sudangrass. In addition, the seeds of pseudosorghum intercepted at the port are easily broken and damaged due to the small cob, glume and other characteristic parts, so the seeds were identified as The main port quarantine work brings more difficulties, therefore, a reliable and fast method to solve this problem is urgently needed in the quarantine wo...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 郭琼霞黄娴黄振虞赟黄可辉沈建国王念武
Owner INSPECTION & QUARANTINE TECH CENT OF FUJIAN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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