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Biological chip of identifying colon canceration degree using microchanging of RNA

A microRNA and biochip technology, applied in the biological field, can solve the problems of lack of specificity and no obvious symptoms in the early manifestations of colon cancer, and achieve the effect of simple and clear results, accurate results, and easy preparation

Inactive Publication Date: 2007-07-18
NANJING UNIV
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AI Technical Summary

Problems solved by technology

[0006] In fact, most of the current clinical cases cannot be diagnosed and treated early. The misdiagnosis rate of colon cancer is quite high. According to some data, the clinical misdiagnosis rate can reach 46.3%. This is because the early manifestations of colon cancer lack specificity and even have no obvious symptoms. , Even if symptoms appear, they are often confused with common intestinal diseases such as intestinal tuberculosis, ulcerative colitis, Crohn's disease, and chronic dysentery, which is the basic cause of misdiagnosis

Method used

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  • Biological chip of identifying colon canceration degree using microchanging of RNA

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Embodiment Construction

[0013] 1. MicroRNA biochip experiment

[0014] In order to screen stable and credible microRNA probes, we have done microRNA biochip experiments, which contain more than 400 probes of mature human microRNAs. SmartArray TM Dots were made on a 75×25mm, chemically modified slide. The samples spotted on the chip also included U6 and tRNA as internal standard, external standard with a length of 30 bases artificially prepared, Hex as positive control for spotting, and 50% DMSO as negative control for hybridization. The entire dot array is divided into 4 sub-arrays, each sub-array has 23 rows and 21 columns, the dot pitch is 185 μm, and the diameter of the dot is about 130 μm, and each probe is repeated three times.

[0015] The specific experimental procedures are as follows: (1) Extraction of total RNA from tissue blocks: Extract total RNA from colon cancer tissues and normal tissues by Trizol method, concentrate RNA by isopropanol precipitation, quantify with spectrophotometer, ...

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Abstract

The present invention belongs to the field of biotechnology and is especially one kind of biochip for identifying the cancerization degree of pathological colon based on the specific change of micro RNA. The present invention solves the technological problem of determining one set of precise and simple probe library to lower the making cost of biochip and simplify the making process of biochip. The technological scheme of the present invention includes screening out micro RNA with obvious and stable change in pathological colon with biochip and quantitative PCR and applying the micro RNA as probe on the biochip to form the biochip for detecting colon cancer pathology. The biochip may be used to diagnose early stage colon cancer.

Description

1. Technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a biochip for identifying the degree of canceration of colonic lesions by specific changes in microribonucleic acid. 2. Background technology [0002] Micro ribonucleic acid, English name microRNA, abbreviated as miRNA, is a kind of non-coding single-stranded small ribonucleic acid molecules with a length of about 19 to 23 nucleotides. They are highly conservative in evolution and widely exist in animal and plant cells. Hundreds of microRNAs have been identified in various biological species such as humans, mice, and rats. MicroRNA can recognize the 3' untranslated sequence of the target mRNA and is not completely complementary to it, thereby inhibiting the translation of the target mRNA. [0003] MicroRNA is related to many normal physiological activities of animals, such as growth and development, fat metabolism and so on. At the same time, microRNAs are also inext...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 张辰宇陈熹郭星张红杰张峻峰项阳巴一王可慧林琳孙国勋张燕朱凌云王军玲戚小强姜晓宏
Owner NANJING UNIV
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