Method for authenticating copy number of target genes in transgenic animal

Abstract

The invention discloses a method for authenticating the copy number of target genes in a transgenic animal. The method provided by the invention comprises the following steps of: taking genome DNA of the transgenic animal as a template; performing fluorescence quantitative PCR by respectively using a target gene primer and a reference gene primer to respectively obtain a fluorescence quantitative PCR result of the target genes and reference genes of the transgenic animal; taking a standard substance with the target genes in a different copy number as the template; performing the fluorescence quantitative PCR by using the target gene primer and the reference gene primer respectively to obtain the fluorescence quantitative PCR result of the target genes and the reference genes in the standard substance; establishing a standard curve relative to the difference values of the cycle indexes of the target genes and the reference genes and the natural logarithm of the copy number of the target genes; and determining the copy number of the target genes in the transgenic animal. The method has the characteristics of simplicity, easy implementation, and accuracy and reliability because the results are consistent by using a conventional Southern blot method.

Description

technical field [0001] The invention relates to a method for identifying the copy number of a target gene in a transgenic animal. Background technique [0002] Transgenic animals (transgenic animals) use laboratory methods to introduce the target genes that people need into their genomes, so that the foreign genes can be integrated with the genes of the animals themselves, and proliferate with cell division, and can be expressed in animals. An animal that is stably passed on to offspring. The foreign structural genes integrated into the animal genome are called transgenes, and the proteins encoded by the transgenes are called transgenic products, which affect animal traits through transgenic products. If the transgene is passed on to offspring, a line or population of transgenic animals is formed. With the continuous deepening of research on transgenic animals and the continuous improvement of experimental techniques, transgenic technology has been more widely used. Almost...

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Problems solved by technology

Facts have proved that it is indeed an effective and accurate method, but it is not difficult to find in the actual operation process that it is time-consuming, laborious, and requires a large number of DNA samples, and even radioactive isotopes (Mason et al., 2003 )

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