Detection method and detection kit for amplifying output signal based on hybridization chain reaction
A technology of hybrid chain reaction and hybrid chain reaction, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, and resistance to vector-borne diseases. And other issues
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Embodiment 1
[0083] Example 1: Two-site hybridization chain reaction amplification detection and detection of novel coronavirus single-stranded nucleotide molecules its kit
specific Embodiment approach
[0085] (1) Target sequence selection:
[0086] The single-stranded nucleic acid molecule to be amplified is selected as the single-stranded ribonucleic acid of the novel coronavirus corresponding to one specific target sequence in the deoxyribonucleotide, the sequence is shown in SEQ ID NO.1, and one non-specific target sequence is selected, The sequence is shown in SEQ ID NO.4,
[0087] (2) Sequence design:
[0088] The probe pair for hybridization chain reaction with the specific target sequence is H1a 1 and H2a 1 , the H1a 1 The sequence is shown in SEQ ID NO.2, the H2a 1 The sequence is shown in SEQ ID NO.3, and the probe pair that hybridizes with the non-specific target sequence is H1b 1 and H2b1, the H1b 1 The sequence is shown in SEQ ID NO.5, the H2b 1 The sequence is shown in SEQID NO.6. The sequence information mentioned in Example 1 is shown in Table 1 below. In the H2b 1 The 5' end of the probe increases a fluorescent group (FAM); the H1a 1 Probes and H1b...
Embodiment 2
[0111] Example 2: Three-site hybridization chain reaction amplification detection and detection of novel coronavirus single-stranded nucleotide molecules its kit
[0112] All the steps in Example 2 are completely consistent with those in Example 1, and will not be described again. The core difference lies in the selection of the target sequence and the design of the probe. specifically:
[0113] One of the specific target sequences is selected, the sequence is shown in SEQ ID NO.1, and the probe pair that hybridizes with the specific target sequence is H1a 1 and H2a 1 , the H1a 1 The sequence is shown in SEQ ID NO.2, the H2a 1 The sequence is shown in SEQ ID NO.3. Two non-specific target sequences are selected, including one non-specific target sequence (1) and one non-specific target sequence (2). The non-specific target sequence (1) The sequence is shown in SEQ ID NO.4, the sequence of the non-specific target sequence (2) is shown in SEQ ID NO.8, and the probe pair f...
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