Process of detecting cell splitting periodic protein gene 6 in tumour susing uracil glucosidase method

Abstract

The present invention relates to uracil glycosidase 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, uracil glycosidase process is adopted for detecting cell division cycle protein gene 6 in tumor. The process has dU to replace dT in synthesizing the first pair of reactants, uracil glycosidase treatment before the second amplification to eliminate the residual pollutant of PCR product, and dUTP to replace dTTP in the second amplification for mixing great amount of dU into the product. The process can eliminate all pollutants of different sources.

Description

technical field [0001] The invention relates to a method for detecting cell division cycle protein gene 6 in tumors by using uracil glucosidase method. 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, it is found that 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 the uracil glucosidase method to detect cell division cycle protein gene 6 in tumors, and makes specific design and experimental verification to improve its specificity and pr...

AI Technical Summary

Problems solved by technology

First, false negative: the most common reasons for false negative results are: Taq DNA polymerase activity is not enough, or the activity is inhibited; primer design is unreasonable, etc.; Generally speaking, the amplified product in the reaction system is real, because the PCR technology is highly sensitive, and a very small amount of target gene contamination will cause a large amount of amplification, resulting in errors in the judgment of the results, so pollution is the main source of false positives in PCR

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