Gene chip for specific detection of bacillus thuringiensis
A technology of Bacillus thuringiensis and Bacillus aureus, which is applied in genetic engineering, plant genetic improvement, and methods based on microorganisms. It can solve the problems of classification and identification, small 16SDNA difference, and poor specificity, and achieve reliable results and provide specificity. Effect
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[0026] Concrete implementation steps of the present invention are as follows:
[0027] Primers and probes were designed by our laboratory and synthesized by Shanghai Sangon Bioengineering Co., Ltd.
[0028] (1) Determination of the target sequence: According to the reported Bacillus thuringiensis 16SDNA sequence and rpoB gene sequence, select the Bacillus thuringiensis rpoB specific gene fragment whose nucleotide sequence is SEQ ID NO: 1, and the nucleotide sequence is SEQ ID NO: 2 The 16S specific gene fragment of Bacillus thuringiensis used as the target sequence for detection.
[0029] (2) Design of primers and probes: After the target sequence is determined, according to the design principles of primers and probes, design primers and probes as follows:
[0030] Bacillus thuringiensis rpoB:
[0031] Primer 1: 5'FAM-GAAGATGCATTACGCAACCTTGAC-3',
[0032] Primer 2: 5'-TGTGAATACCAAGGTATCTTGCTG-3',
[0033] 5'NH 3 — TTTTTTTTTTTT ACTACGTGTACCACACGGTGGTGGCG — 3'.
[0034] Ba...
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