Method for carrying out rapid amplification of whole blood genome

Abstract

The invention discloses a method for carrying out rapid amplification of a whole blood genome applied to the field of analysis and research of biomedical molecules. The method for carrying out rapid amplification of the whole blood genome is achieved by a nucleic releaser and polymerase chain reaction (PCR) reaction liquid matched with the nucleic releaser. The method comprises the following main steps: adding 5mu l of nucleic releaser to a PCR reaction tube; mixing the mixed solution from the former step with 5mu l of blood, standing at room temperature for 5-10 minutes, and then directly adding 40mu l of prepared PCR reaction liquid to the mixed liquor; finally adding to the machine to amplify, wherein the nucleic releaser comprises 0.4-2% of NaOH, 0.02-2% of Chaps, 0.5-20% of (NH4)2SO4, and 0.1-5% of formamide, and the ratio is 100%; the PCR reaction liquid comprises 30mmol / L of Tris-HCl (PH7.5), 10mmol / L of (NH4)2SO4, 30mmol / L of KCl, and 4.5mmol / L of MgCl2. By adopting the method, a complicated blood nucleic acid purification process is emitted, a 'one-room' operation of blood nucleic acid extraction and amplification is achieved, nucleic acid loss caused in the nucleic acid extraction process is avoided, the operation is simple and easy to grasp, the operation method is convenient and fast, the cost is low and the amplification efficiency is high.

Description

technical field [0001] The invention relates to a method for rapidly amplifying whole blood genome in the field of biomedical molecular analysis research. Background technique [0002] In recent years, the application of PCR technology to analyze human or animal genetic background has become an important technical analysis method. In the process of genetic background analysis, the main sample material is blood. Since blood contains a large amount of DNA polymerase inhibitors, such as heme, miscellaneous protein, fat, etc., PCR method is used to directly amplify the nucleic acid in blood. Totally impossible. In order to complete the nucleic acid amplification of blood samples, the nucleic acid substances must be purified from the blood. The classic blood nucleic acid extraction method mainly includes several processes. 1. The erythrocytes are ruptured by the hypotonic fluid, and the hemoglobin substance is released, and then centrifuged and washed to remove the hemoglobin....

AI Technical Summary

Problems solved by technology

However, due to the use of defective Taq enzymes in this methodology, the fidelity of the PCR amplification process guided by Taq enzymes is low, which often leads to non-specific amplification, resulting in false positive results.

At the same time, due to the use of these special Taq enzymes, the cost of detection and amplification is very high, which is 20-30 times that of ordinary Taq enzyme amplification

This also makes it very difficult to promote this method due to the high cost

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