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Primer, probe, kit and method for detecting transgenic rice strain T1c-19

A technology of transgenic rice, t1c-19, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., to achieve the effect of solving the pollution after PCR

Inactive Publication Date: 2013-04-24
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0016] At present, there are few reports at home and abroad that can quickly, simply, specifically and sensitively detect the transgenic rice line T1c-19 or its components

Method used

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  • Primer, probe, kit and method for detecting transgenic rice strain T1c-19
  • Primer, probe, kit and method for detecting transgenic rice strain T1c-19
  • Primer, probe, kit and method for detecting transgenic rice strain T1c-19

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Specificity Test

[0042] The present embodiment tests the specificity of oligonucleotide primers and probes of the present invention, wherein specific primer OrylC-F / R is used, that is, the base sequences of the oligonucleotide primers used are SEQ ID NO:1 and SEQ ID NO: 2, the base sequence of the probe is Ory1C-P (SEQ ID NO: 3), a fluorescent quenching group BHQ1 is connected to the 3'-end of the probe, and a fluorescent quencher is connected to the 5'-end Reporting group FAM.

[0043] In this example, samples of the transgenic rice line T1c-19 and samples of common rice lines with similar relatives were tested, including: Kangyou 97, Kefeng 6, Kefeng 8, Huahui 1, Kemodao KMD1, LL62, LL601, T2a-1, Batunmi, K105, Minghui 63, Peiza 35, Jindao 9618, Wandao 181 and Chunyou 59. In addition, a sample without DNA was used as a blank control.

[0044] Main testing instruments :

[0045] Micropipette (1000uL, 100uL, 10uL or 20uL, 2.5uL; Eppendorf); fluorescen...

Embodiment 2

[0080] Embodiment 2: sensitivity test

[0081]The sensitivity of the oligonucleotide primers and probes of the present invention, wherein the specific primer OrylC-F / R is used, the base sequence of the oligonucleotide primer used is SEQ ID NO:1 and SEQ ID NO:2, The base sequence of the probe is Ory1C-P (SEQ ID NO: 3), a fluorescent quenching group BHQ1 is connected to the 3'-end of the probe, and a fluorescent reporter group FAM is connected to the 5'-end.

[0082] The main detection instruments and reagents used in this example are as shown in Example 1, but transgenic rice lines T1c-19 with different relative mass fractions (2%, 0.2%, 0.01%, w / w) were used in the test The DNA of the sample was carried out in duplicate at each concentration, so as to detect the sensitivity and detection limit of the transgenic rice line T1c-19 sample. The experimental results are shown in figure 2 middle.

[0083] Depend on figure 2 It can be seen that the sensitivity of the oligonucleo...

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Abstract

The invention relates to a primer, a probe, a kit containing the primer and the kit for detecting a transgenic rice strain T1c-19 or the components of the transgenic rice strain T1c-19 by using a real-time PCR (Polymerase Chain Reaction) method, and a real-time PCR detection method for detecting the transgenic rice strain T1c-19 or the components of the transgenic rice strain T1c-19 by using the primer and the probe.

Description

technical field [0001] The invention belongs to the field of biotechnology. More specifically, the present invention relates to primers and probes for detecting transgenic rice line T1c-19 or its components by real-time PCR method, a kit comprising the primers and probes, and detection of Real-time PCR detection method for components or components of transgenic rice line T1c-19. Background technique [0002] Genetically modified organisms refer to the use of biotechnology to transfer foreign genes to other species to modify their genetic characteristics, so as to obtain the traits and nutritional qualities required by human beings. Foods that are processed from genetically modified organisms or their raw materials are genetically modified foods. Since the birth of the first genetically modified food (transgenic tomato) in the United States in 1994, genetically modified organisms have widely entered the food field. [0003] Breeding of transgenic insect-resistant rice [...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 黄文胜陈颖邓婷婷付凯
Owner CHINESE ACAD OF INSPECTION & QUARANTINE