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PCR method for quantitative detection of abi1-tsv-11 and its primers
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A technology of ABI1-TSV-11 and internal primers, applied in biochemical equipment and methods, microbe determination/inspection, DNA/RNA fragments, etc.
Active Publication Date: 2022-04-12
PEOPLES HOSPITAL PEKING UNIV
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[0004] At present, there is still a lack of rapid, effective and accurate quantitative detection method of ABI1-TSV-11 at the nucleic acid level in clinical practice, and the application of nested PCR two-step method for quantitative detection of colon cancer cancer-derived genes has not been seen at home and abroad. ABI1 The No. 11 transcript variant of ABI1-TSV-11 was reported
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Embodiment 1
[0062] Embodiment 1, design and synthesis of nested PCR primers
[0063] 1. Design and synthesis of ABI1-TSV-11 nested PCR detection primers
[0064] Abelson interactor 1 (ABI1) gene ABI1 After transcription, 18 transcript variants (ABI1-TSV-1 mRNA, ABI1-TSV-2 mRNA, ABI1-TSV-3 mRNA, ABI1-TSV-4 mRNA, ABI1-TSV-5 mRNA, ABI1 -TSV-6 mRNA, ABI1-TSV-7 mRNA, ABI1-TSV-8 mRNA, ABI1-TSV-9 mRNA, ABI1-TSV-10 mRNA, ABI1-TSV-11 mRNA, ABI1-TSV-12 mRNA, ABI1- TSV-13 mRNA, ABI1-TSV-14 mRNA, ABI1-TSV-15 mRNA, ABI1-TSV-16 mRNA, ABI1-TSV-17 mRNA, and ABI1-TSV-18 mRNA). Using NCBI published ABI1 The mRNA nucleic acid sequence conservation region (Exon 3 and Exon 13) of the 18 TSVs molecules of the gene was designed for the first round of nested PCR outer primers: ABI1-TSVs (Abelson Interacting Protein 1 Transcriptional Variant) universal primer (outer primer): Primer ABI1-F (corresponding to Exon 3) and primer ABI1-R (corresponding to Exon13).
[0065] Using the Exon-Exon junction (Exon-Exon J...
[0071]In this example, the established ABI1-TSV-11 overexpression cell lines (SW480 and LoVo) and 4 pairs of colorectal cancer (Colorectal Cancer, CRC) clinical tissue samples (including cancer and paracancerous tissues) were used as samples to be tested. Nested-PCR (Nested-PCR) relative quantitative detection experiment to confirm the application value of this detection technology in experimental research and clinical research.
[0072] 1. Samples to be tested:
[0073] (1) Cell samples: ABI1-TSV-11 overexpression cell lines (SW480 and LoVo) were described in the following literatures: Zhang Y, Zhong Z, Li M, Chen J, Lin T, Sun J, Wang D, Mu Q, Su H, Wu N, LiuA, Yu Y, Zhang M, Liu Y, Guo J, Yu W. The roles and prognostic significance of ABI1-TSV-11 expression in patients with left-sided colorectal cancer. SciRep. 2021 May 24;11 (1):10734. doi: 10.1038 / s41598-021-90220-8. PMID: 34031495; PMCID: PMC8144562.
[00...
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Abstract
The invention discloses a PCR method for quantitatively detecting ABI1-TSV-11 and the primers used therefor. Specifically disclosed is a nested PCR primer set for detecting ABI1-TSV-11 and an application thereof. The present invention is based on existing 18 kinds ABI1 The molecular structure of gene TSVs and the molecular structure specificity of ABI1‑TSV‑11 were designed, related primers and PCR reaction conditions were designed, and a set of rapid, effective, stable and accurate ABI1‑TSV‑11 nested PCR quantitative detection technology was established , this technology has the characteristics of good specificity and high sensitivity, can effectively avoid false positive amplification, and break through the bottleneck of specific detection of ABI1-TSV-11 at the nucleic acid level, as a new molecular biology technology in the knot colorectal canceroncogene ABI1 It has potential application value in the detection of transcript variants.
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