Multivalent probes having single nucleotide resolution
A nucleic acid probe and single-stranded oligonucleotide technology, which is used in the determination/inspection of microorganisms, biochemical equipment and methods, etc., and can solve problems such as lack of specificity and inability to detect single-base substitutions.
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Embodiment 1
[0200] Example 1: Using Polymer Strand Pair / Partially Double-Stranded Probes with NanoString Technologies ® SNP detection experiments of existing DV2 reporter probes
[0201] An experiment was performed to detect the V600E single nucleotide polymorphism (SNP) of the BRAF gene.
[0202] Figure 18A The three partially double-stranded probes used in this example are shown: the first probe to detect the wild-type target and the two probes (ml and m2) to detect the two mutant forms of the target. In this experiment, using NanoStringTechnologies ® DV2 system. The probes included target binding regions of 10 nucleotides and 8 nucleotides in length ("10+8"), wherein m1 and m2 mutations were detected by the 8 nucleotide target binding region.
[0203] Synthetic targets (corresponding to BRAF exon 15) for the three probes are shown in Figure 18B middle. In this experiment, the target of the first target binding domain and the target of the second target binding domain are contig...
Embodiment 2
[0215] Example 2: Using Polymer Strand Pairs / Partially Double-Stranded Probes with NanoString Technologies in the Hotspot Region ® SNV Detection Experiments of Existing DV2 Reporter Probes
[0216]Experiments to detect single nucleotide variants (SNVs) in the KRAS exon 2 hotspot were performed.
[0217] Figure 26 A sketch of a partially double-stranded probe similar to that used in the examples herein is shown. In this experiment, NanoString Technologies ® DV2 system. Note that any of the probes, probe pairs, or compositions shown in Figures 1-15 may be substituted for this example, or any of the examples disclosed herein. Figure 26 Probes shown. exist Figure 26 , single nucleotides on the target sequence (gray) can be detected with existing NanoString Reporter oligos (green) using a two-armed linker probe called "Probe A" (blue). Linker probes consist of two oligomers hybridized together by a stem sequence. In these experiments, two oligos hybridize to nearby regi...
Embodiment 3
[0223] Example 3: Using Polymer Strand Pairs / Partially Double-Stranded Probes with NanoString Technologies ® Deletion detection experiments for existing DV2 reporter probes
[0224] Experiments to detect deletions in EGFR exon 19 were performed.
[0225] In this experiment, NanoString Technologies ® DV2 system with Figure 26 The structure of the dual-arm probe is shown. Note that for this example, or any of the examples disclosed herein, any of the probes, probe pairs, or compositions shown in Figures 1-15 may be substituted for Figure 26 Probes shown. tested against Figure 30 Shown are reference sequences and probes specific for the three targets containing the deletion. Each probe includes two target binding regions and is 18-21 nucleotides in length. The sequence and position of each probe are shown in Figure 31A -D ( Figure 31A show the entire sequence and Figure 31B -D display each Figure 31A one-third of the sequence). Overall, the probe library conta...
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