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Detection method for genetically modified wheat B73-6-1

A technology of transgenic wheat and detection method, which is applied in the field of identification of transgenic wheat strains, can solve the problems of unreliable screening effect and achieve high sensitivity and strong specificity

Inactive Publication Date: 2013-06-26
曹际娟 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Selectable marker genes (such as neomycin phosphotransferase gene, chloramphenicol acetyltransferase gene, hygromycin transferase gene, etc.) and reporter genes (such as B2 glucuronidase gene and luciferase gene, etc.) have been widely For screening of transformed plants and tissues, but in some cases, this selection is unreliable

Method used

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  • Detection method for genetically modified wheat B73-6-1
  • Detection method for genetically modified wheat B73-6-1
  • Detection method for genetically modified wheat B73-6-1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1. Acquisition of complete sequences of transgenic wheat-specific gene components A, B, and C and corresponding border sequences

[0024] 1. Genomic DNA extraction from transgenic samples

[0025] The transgenic wheat B72-8-11b, transgenic wheat B73-6-1 and transgenic wheat B102-1-2 were respectively selected and marked as sample A, sample B and sample C, respectively. Each sample was pulverized and ground into a fine powder in liquid nitrogen, and then genomic DNA was extracted using DNA Extraction Kit for GMO Detection Ver.2.0 (TaKaRa Code No. D9093).

[0026] Detect the quality of the DNA sample by 1% agarose gel electrophoresis, and use an ultramicro spectrophotometer (Nanodrop) to detect the quality and concentration of the genomic DNA of the sample. The test results show that the quality of the genomic DNA meets the requirements of subsequent experiments, and samples A, B and C The average genomic DNA concentrations were 162.69ng / μL, 125.80ng / μL, and 203....

Embodiment 2

[0101] Embodiment 2. Applied to the detection of actual samples

[0102] 1. Sample preparation:

[0103] Select 10 wheat samples to be tested (as shown in Table 4), grind them into fine powder in liquid nitrogen, and use DNA Extraction Kit for GMO Detection Ver.2.0 (TaKaRa Code No.D9093) for genomic DNA extraction. Then, the quality of genomic DNA was detected by agarose gel electrophoresis, and the quality and concentration of genomic DNA were detected by Nanodrop to determine that the quality and concentration of genomic DNA of the sample met the requirements of subsequent experiments.

[0104] 2. The 10 samples were tested according to method A, method B and method C established in Example 1, and the results are shown in Table 4, wherein "+" indicates that the amplification result is positive, and "-" indicates that the amplification result is negative.

[0105] From the test results in Table 4, it can be seen that

[0106] Method A can accurately, sensitively and specif...

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Abstract

The invention relates to a detection method for genetically modified wheat B73-6-1, namely detecting a genetic component of which the sequence is SEQ ID NO: 12 by a PCR (polymerase chain reaction) method. The genetic component comprises edge sequences of which the sequences are SEQ ID NO: 15 and SEQ ID NO: 16. By the method, variety identification of the genetically modified wheat B73-6-1 can be quickly and sensitively realized. The method has high specificity and high sensitivity and is fast and accurate.

Description

technical field [0001] The invention relates to a method for identifying transgenic wheat strains. Background technique [0002] Transgenic wheat is the use of genetic engineering technology to introduce foreign genes into wheat genes, and the wheat that can express the target gene is obtained after screening. In gene transfer and expression studies, molecular testing of transformed seedlings is required to weed out non-transformed plants. Selectable marker genes (such as neomycin phosphotransferase gene, chloramphenicol acetyltransferase gene, hygromycin transferase gene, etc.) and reporter genes (such as B2 glucuronidase gene and luciferase gene, etc.) have been widely Used for screening of transformed plants and tissues, but in some cases, this selection is unreliable. PCR is one of the commonly used techniques for detection for this purpose, which has the advantages of sensitivity and rapidity. The application of this technology is to find appropriate and highly speci...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 曹际娟徐君怡肖珊珊
Owner 曹际娟