Nucleic acid adapter used for detecting human high-metastasis colon cancer cell strain LoVo and detection kit

A nucleic acid aptamer and colon cancer cell technology, which is applied in the field of bioengineering to achieve the effects of small molecular weight, easy labeling and low preparation cost

Active Publication Date: 2016-03-09
HUNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the lack of corresponding technology to obtain highly specific and sensitive molecular probes for ...

Method used

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  • Nucleic acid adapter used for detecting human high-metastasis colon cancer cell strain LoVo and detection kit
  • Nucleic acid adapter used for detecting human high-metastasis colon cancer cell strain LoVo and detection kit
  • Nucleic acid adapter used for detecting human high-metastasis colon cancer cell strain LoVo and detection kit

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Embodiment 1

[0041] A nucleic acid aptamer that can be used to detect the human highly metastatic colon cancer cell line LoVo was screened by the following method:

[0042] 1. Synthesize the random single-stranded DNA library and primers shown in the following sequences:

[0043]Random library RL40: 5'-CCTGAACCTGATGCCAACCT(N40)AGTAGCGAGCGTGTAGTGTG-3';

[0044] 5' primer: 5'-FAM-CCTGAACCTGATGCCAACCT-3';

[0045] 3' primer: 5'-Biotin-CACACTACACGCTCGCTACT-3'.

[0046] 2. Cell-SELEX screening to obtain LoVo-specific nucleic acid aptamer library (positive screening):

[0047] 2.1. Cell pretreatment: Add 10% calf serum to culture medium 1640 to culture LoVo cells of human highly metastatic colon cancer cell line until 90% of the bottom of the bottle is covered, remove the medium, wash with PBS buffer, and use enzyme-free digestion solution (purchased from Beijing Pulilai Gene Technology Co., Ltd.) The cells were processed into suspension cells, transferred to centrifuge tubes, and centrifuged...

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Abstract

The invention discloses a nucleic acid adapter used for detecting a human high-metastasis colon cancer cell strain LoVo and a detection kit. The nucleic acid adapter is one of a nucleic acid adapter J1, a nucleic acid adapter J3, a nucleic acid adapter J4 and a nucleic acid adapter J7; the nucleotide sequence of the nucleic acid adapter J1 is a DNA fragment shown as SEQ ID NO.1; the nucleotide sequence of the nucleic acid adapter J3 is a DNA fragment shown as SEQ ID NO.2; the nucleotide sequence of the nucleic acid adapter J4 is a DNA fragment shown as SEQ ID NO.3; the nucleotide sequence of the nucleic acid adapter J7 is a DNA fragment shown as SEQ ID NO.4. The nucleic acid adapter is low in immunogenicity, small in molecular weight, easy to store and label and capable of serving as the detection kit to detect the human high-metastasis colon cancer cell strain LoVo.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a nucleic acid aptamer for detecting the LoVo human highly metastatic colon cancer cell line, a screening method and application thereof. Background technique [0002] Tumor metastasis is the most important cause of cancer death, and more than 50% of patients with colon cancer have tumor metastasis. The prognosis of these patients with tumor metastasis is extremely poor, and the five-year survival rate is only about 6%. Therefore, the development of specific methods to detect and treat colon cancer metastases is crucial to improve the survival of these patients. However, due to the lack of corresponding technology to obtain highly specific and sensitive molecular probes for colon cancer tumor metastasis, the diagnosis and treatment of highly metastatic colon cancer are still facing great challenges. In recent years, with the advancement of imaging and surgical techniques,...

Claims

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Application Information

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IPC IPC(8): C12N15/115G01N33/574
CPCC12N15/115C12N2310/16G01N33/57419C12N2310/315C12N2310/3181
Inventor 王柯敏袁宝银郭秋平羊小海何晓晓蒋晓春
Owner HUNAN UNIV
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