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Spire tetrahedral DNA nano-structural probe and telomerase electrochemical detection

A nanostructure and tetrahedral technology, applied in the direction of recombinant DNA technology, material electrochemical variables, DNA / RNA fragments, etc., can solve the problems of reduced telomerase activity and low sensitivity

Inactive Publication Date: 2015-01-07
SHANGHAI INST OF MEASUREMENT & TESTING TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem solved by the present invention is to overcome the defects of low sensitivity and reduced telomerase activity due to the surface crowding effect in the existing telomerase detection test, and provide a double-stranded spire tetrahedral DNA nanostructure probe and its preparation method , greatly improve the signal-to-noise ratio and sensitivity of telomerase detection, and have a wider detection range of cell number

Method used

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  • Spire tetrahedral DNA nano-structural probe and telomerase electrochemical detection
  • Spire tetrahedral DNA nano-structural probe and telomerase electrochemical detection
  • Spire tetrahedral DNA nano-structural probe and telomerase electrochemical detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] 1 Construction of double-stranded spire tetrahedral DNA nanostructured probe (STTS), single-stranded DNA probe (ssP) without tetrahedral structure and three-dimensional DNA nanostructured probe (TSP) with ordinary tetrahedral structure.

[0045] Experimental Materials:

[0046] STTS: STTS-A (93bp, molecular weight 28012.0, ssDNA), TSP-B (55bp, molecular weight 17018.0, 5' end modified sulfhydryl ssDNA), TSP-C (55bp, molecular weight 16898.0, 5' end modified sulfhydryl ssDNA) and TSP- D (55bp, molecular weight 16877.0, ssDNA modified with sulfhydryl group at the 5' end) and complementary strand Spire-Complementary DNA (SC-DNA, 20bp, molecular weight 6176.0, ssDNA), the nucleotide sequences of which are shown in the sequence table as SEQ ID No.1 and SEQ ID No.1, respectively Shown in ID No.4, SEQ ID No.5, SEQ ID No.6 and SEQ ID No.7.

[0047] ssP: ss-primer (28bp, molecular weight 8449.0, ssDNA with sulfhydryl group modified at the 5' end), the nucleotide sequence is sho...

Embodiment 2

[0072] 1 Construction of double-stranded spire tetrahedral DNA nanostructure probe (STTS)

[0073] Experimental Materials:

[0074] STTS-A (93bp, molecular weight 28012.0, ssDNA), TSP-B (55bp, molecular weight 17018.0, ssDNA modified with thiol at the 5' end), TSP-C (55bp, molecular weight 16898.0, ssDNA with thiol modified at the 5' end) and TSP-D ( 55bp, molecular weight 16877.0, 5' end modified sulfhydryl ssDNA) and complementary strand Spire-Complementary DNA (SC-DNA, 20bp, molecular weight 6176.0, ssDNA), the nucleotide sequences are respectively as SEQ ID No.1 and SEQ ID No in the sequence table .4, shown in SEQ ID No.5, SEQ ID No.6 and SEQ ID No.7.

[0075] The single-stranded DNAs shown in SEQ ID No.1, 4-7 in the sequence listing were all synthesized by Life Technology Biological Co., Ltd.

[0076] Experimental steps:

[0077] Take a gold electrode chip (SC1000-16X, GENE fluidics) and soak it in isopropanol for 1 min, then use ultrapure water to sonicate for 10 s, a...

Embodiment 3

[0093] 1 Construction of double-stranded spire tetrahedral DNA nanostructure probe (STTS)

[0094] Experimental Materials:

[0095]STTS-A (93bp, molecular weight 28012.0, ssDNA), TSP-B (55bp, molecular weight 17018.0, ssDNA modified with thiol at the 5' end), TSP-C (55bp, molecular weight 16898.0, ssDNA with thiol modified at the 5' end) and TSP-D ( 55bp, molecular weight 16877.0, 5' end modified sulfhydryl ssDNA) and complementary strand Spire-Complementary DNA (SC-DNA, 20bp, molecular weight 6176.0, ssDNA), the nucleotide sequences are respectively as SEQ ID No.1 and SEQ ID No in the sequence table .4, shown in SEQ ID No.5, SEQ ID No.6 and SEQ ID No.7.

[0096] The single-stranded DNAs shown in SEQ ID No.1, 4-7 in the sequence listing were all synthesized by Life Technology Biological Co., Ltd.

[0097] Experimental steps:

[0098] Take a gold electrode chip (SC1000-16X, GENE fluidics) and soak it in isopropanol for 1 min, then use ultrapure water to sonicate for 10 s, an...

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Abstract

The invention discloses a double-stranded spire tetrahedral DNA nano-structural probe (STTS) as well as a preparation method and an application of the STTS in electrochemical detection of the activity of a telomerase. The STTS comprises a single-stranded probe STTS-A, a single-stranded probe B, a single-stranded probe C, a single-stranded probe D and a single-stranded probe SC-DNA. The invention also discloses a method for electrochemical detection of the activity of the telomerase by virtue of the STTS and the application of the STTS in electrochemical detection of the activity of the telomerase. By adopting the method for electrochemical detection of the activity of the telomerase, the signal to noise ratio of detected signals is more than twice higher than that of the detected signals detected the ordinary TSP (Test Sphere Probe) detection, the sensitivity is high, the specificity is strong and the application range is wide.

Description

technical field [0001] The invention specifically relates to a double-stranded spire tetrahedron DNA nanostructure probe, a preparation method thereof and an application in electrochemical detection of telomerase activity. Background technique [0002] Telomeres are repetitive segments of DNA located at the ends of chromosomes in eukaryotic cells that maintain chromosome stability and integrity. Telomeres are composed of telomere DNA and telomere-related proteins, and are special structures located at the ends of chromosomes in eukaryotic cells. Telomerase is composed of RNA subunit (human telomerase RNA, hTR), telomerase catalytic subunit with reverse transcriptase activity (human telomerase reverse transcriptase, hTERT) and telomerase associated protein (telomere associated protein, TEP). Partial protein complex. Telomerase is a reverse transcriptase that uses its own RNA as a template to specifically synthesize the telomere DNA repeat sequence at the 3' end of the chrom...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/48C12Q1/28C12Q1/26G01N27/26
Inventor 刘刚李妍闻艳丽梁文许丽王乐乐
Owner SHANGHAI INST OF MEASUREMENT & TESTING TECH
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