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Detection kit and method of common transgenic soybean strains

A technology of genetically modified soybeans and detection kits is applied in biochemical equipment and methods, and microbial determination/inspection. The effect of improving sensitivity and specificity

Active Publication Date: 2018-10-19
SICHUAN HUAHAN TRIO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Qualitative PCR detection generally uses gel electrophoresis analysis, which has certain limitations, and can only detect DNA bands above 10ng, so it cannot be detected when the yield of the target fragment is low
Real-time fluorescence quantitative PCR detects the amplification amount of PCR products in real time by detecting the strength of the fluorescent signal, and can quantitatively analyze the genetically modified components, but it can only do single-fold or up to 3-4 multiples, and cannot increase its throughput.

Method used

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  • Detection kit and method of common transgenic soybean strains
  • Detection kit and method of common transgenic soybean strains
  • Detection kit and method of common transgenic soybean strains

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] The detection kit of the common transgenic soybean strain provided in this example includes a nucleic acid combination and a membrane chip, wherein the nucleic acid combination includes 13 primer pairs and positive oligonucleotide single-stranded DNA, and the base of the upstream primer and the downstream primer of each primer pair The base sequence (5'-3') is as follows:

[0104] The first primer pair used to detect the GTS40-3-2 strain:

[0105] It includes the upstream primer shown in SEQ ID NO.1 and the downstream primer shown in SEQ ID NO.2;

[0106] Second primer pair for detection of DP356043 strain:

[0107] It includes the upstream primer shown in SEQ ID NO.3 and the downstream primer shown in SEQ ID NO.4;

[0108] The third primer pair used to detect the DP305423 strain:

[0109] It includes the upstream primer shown in SEQ ID NO.5 and the downstream primer shown in SEQ ID NO.6;

[0110] The fourth primer pair for detecting the A2704-12 strain:

[0111] I...

Embodiment 2

[0158] Adopt the detection kit of embodiment 1 to detect the method for transgenic soybean line, specifically as follows:

[0159] 2.1 Extract the genomic DNA of the sample to be tested, and dilute the DNA solution to the same mass concentration (about 50 ng / μL) with a micro-nucleic acid analyzer to obtain the DNA template of the sample to be tested.

[0160] 2.2 Multiplex PCR system and conditions

[0161] The reaction system of PCR amplification:

[0162] 10×PCR Buffer (with Mg 2+ , containing UNG enzyme): 5 μL;

[0163] dNTP (2.5mM each): 5μL;

[0164] Biotin-labeled downstream primers (20 μM): 1.5 μL; upstream primers (20 μM): 1 μL; instructions: 13 primer pairs (provided in Example 1) were added, each primer pair had 1.5 μL of downstream primers, and 1.5 μL of upstream primers 1 μL;

[0165] DNA template of the sample to be tested (50ng / μL): 2μL;

[0166] Positive oligonucleotide DNA ((20μM)): 0.01μL;

[0167] ddH 2 O to a total volume of 50 μL. Perform multiple ...

Embodiment 3

[0197] Take GTS40-3-2, DP356043, DP305423, A2704-12, A5547-127, FG72, CV127, MON87701, MON87705, MON87708, MON87769, MON89788 transgenic soybean line DNA as templates respectively, using the kit of Example 1, according to the implementation The method of example 2 detects, and the result is as follows figure 2 shown, from figure 2 It can be seen from the results that each transgenic soybean line was detected, and there was no cross-reaction among each other, indicating that the kit in Example 1 had better specificity.

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Abstract

The invention discloses a detection kit and method of common transgenic soybean strains and relates to the technical field of transgenic detection. The detection kit comprises a combination of one ormore of the following primer pairs: SEQ ID NO.1-2, SEQ ID NO.3-4, SEQ ID NO.5-6, SEQ ID NO.7-8, SEQ ID NO.9-10, SEQ ID NO.11-12, SEQ ID NO.13-14, SEQ ID NO.15-16, SEQ ID NO.17-18, SEQ ID NO.19-20, SEQID NO.21-22 and SEQ ID NO.23-24. By adopting the kit to detect transgenic soybean strains, the specificity and the sensitivity are higher, and the detection result is accurate and reliable.

Description

technical field [0001] The invention relates to the technical field of transgenic detection, in particular to a detection kit and method for common transgenic soybean strains. Background technique [0002] Soybean is rich in protein and fat. It is an important oil crop and a high-protein grain and feed crop. It is also a high-quality raw material for various processing industries such as food and feed. In recent years, the number of imported soybeans in my country has increased year by year. With the rapid development of commercial planting of genetically modified crops around the world, the safety of genetically modified foods has become more and more controversial, and there is no conclusion yet. Since the end of 1993, my country has officially promulgated the "Genetic Engineering Safety Management Method", which has made clear regulations on the genetic engineering biological experiments carried out in the territory. The State Council also promulgated the "Regulations o...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6834
CPCC12Q1/6834C12Q1/6895C12Q2600/16C12Q2600/166C12Q2531/113C12Q2537/143C12Q2545/101
Inventor 黄盈韩嘉钰陈浩东
Owner SICHUAN HUAHAN TRIO BIOTECH CO LTD
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