Method for quick volume-production of PCR mould plate

Abstract

The invention discloses a method for preparing a PCR template rapidly in a large batch manner, belongs to the technology field of plant molecule detection, and in particular relates to thermal cracking of plant tissues in lysate in order to realize the neutralization with the lysate. The method adopts the technical proposal that trace plant leaves are put in the lysate with certain volume for thermal cracking and neutralization; reaction mixed solution can be used as the template of PCR reaction directly; the PCR reaction is stable and the repeatability is strong under the circumstance of adding with proper polymerase stabilizing agent. The method is particularly suitable for the rapid preparation of the PCR reaction template in the large batch manner, greatly increases the efficiency of the molecule detection and the genetic analysis, and lowers the cost extremely markedly.

Description

technical field [0001] The invention relates to a method for preparing a PCR template, in particular to a method for rapidly preparing a PCR template in large quantities and a reagent formula. After the plant tissue is lysed by the lysate and neutralized by the neutralizer, it is directly used as a PCR reaction template for PCR amplification analysis, providing a template for large-scale genetic analysis of plant genomes. Background technique [0002] With the rapid development of molecular biology, the research field of biotechnology has gradually expanded to all aspects of biological science. The study of nucleic acid level is an important content of molecular biology research. PCR (polymerase chain reaction) technology is the main tool for the study of nucleic acid. Playing an increasingly important role in the field of molecular biology. The premise of using PCR technology to study nucleic acid is to prepare a template for PCR reaction, that is, to release genetic mater...

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Problems solved by technology

[0003] At present, there are mainly CTAB (Wang Li, Qiao Aimin, Sun Yiming, etc.) methods for extracting plant genomic DNA more conventionally. Genomic DNA extraction of Chinese cabbage and optimization of RAPD reaction system [J]. Journal of Southwest Normal University, 2006, 31 (2): 124-128) and SDS (Nie Zhensu, Lai Zhongxiong, Pan Dongming, etc. Olive genome DNA extraction and RAPD amplification conditions optimization [J]. Subtropical Agricultural Research, 2005, 1 (2): 6-8) method, this The two methods have their own advantages and disadvantages, but both require low-temperature grinding, high-temperature cracking, residue separation, removal of impurities such as protein, RNA, and salts, precipitation of DNA, and centrifugation of plant tissues. The procedures are complex, cumbersome, and time-consuming. , high cost, more plant materials are needed, and organic reagents harmful to human body such as phenol and chloroform are used in the extraction process

In addition, a lot of medicines are needed, especially when DNA samples need to be extracted from large quantities of materials, because many utensils have to be reused in the process of grinding and extracting DNA, which often has a serious impact on the PCR reaction, which is highly sensitive to contamination

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