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Process of detecting cell splitting periodic protein gene 6 in tumour using heat starting polyases chain reaction

A cell division and cycle protein technology, applied in the field of detection of cell division cycle protein gene 6 in tumors, can solve the problems of cumbersome methods and easy pollution, and achieve the effects of ensuring reliability, preventing infection, and improving detection sensitivity and specificity

Inactive Publication Date: 2007-07-18
张曼
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AI Technical Summary

Problems solved by technology

Delayed addition of TaqDNA polymerase means that when the reaction system reaches 90 degrees, PAUSE, keep the temperature above 70 degrees, and add polymerase manually, but this method is too cumbersome, especially for high-throughput applications, and it is easy to cause pollution

Method used

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  • Process of detecting cell splitting periodic protein gene 6 in tumour using heat starting polyases chain reaction
  • Process of detecting cell splitting periodic protein gene 6 in tumour using heat starting polyases chain reaction

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Embodiment Construction

[0011] 1. Randomly select cases

[0012] 30 cases of bladder cancer were diagnosed, including 6 cases of initial onset and 24 cases of recurrence; 26 cases of men aged 36 to 84; 4 cases of women aged 60 to 73.

[0013] 25 cases of other urinary system diseases were diagnosed, including 18 cases of males aged 28-77; 7 cases of females aged 27-69; causes: 4 cases of bladder stones; 4 cases of kidney stones; 10 cases of urinary tract infection; 2 cases of kidney cancer; epididymitis 1 case; 2 cases of benign prostatic hyperplasia; 2 cases of urethral stricture. 5 male healthy volunteers aged 29-31.

[0014] 2. Preparation of Urinary Exfoliated Cells

[0015] Take 150-200ml of random urine after the first urine in the morning, divide it into several treated glass centrifuge tubes, centrifuge at 4°C and 380g for 10 minutes, discard the supernatant, collect the precipitate in a centrifuge tube, and wash with normal saline The pellet was washed twice, and the pellet was divided in...

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Abstract

The present invention relates to hot starting PCR process for detecting cell division cycle protein gene 6 in tumor. In the process of DNA extraction, RNA extraction, protein extraction, immunohistochemistry, reverse transcription (RT), PCR, Northern hybridizing, in-situ hybridizing and capillary electrophoresis, heat resistant DNA polymerase (Taq DNA polymerase) is adopted for hot starting PCR process of detecting cell division cycle protein gene 6 in tumor. The present invention has raised detection sensitivity and specificity, high reliability of clinical judgment of tumor cell course and prognosis, and important significance in preventing infection.

Description

technical field [0001] The invention relates to a method for detecting cell division cycle protein gene 6 in tumors by using hot start polymerase chain reaction. Background technique [0002] With the development of molecular biology techniques, gene diagnosis and gene therapy are more and more widely used in clinic, such as the development and application of polymerase chain reaction (polymerase chain reaction, PCR). However, due to the characteristics of PCR technology itself, false positives and false negatives are prone to occur in clinical applications. In order to avoid these errors, many specific changes, designs, analyzes and experimental verifications should be made according to the characteristics of different nucleic acid fragments in the actual application of PCR. . This patent uses hot-start polymerase chain reaction to detect cell division cycle protein gene 6 in tumors. [0003] Cell division cyclin gene 6 (cdc6) is the primary factor for the initiation and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 张曼
Owner 张曼
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