Two-step PCR technology

A technical and specific technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of time-consuming and other problems, and achieve the effect of short time-consuming, wide adaptability and high cost

Pending Publication Date: 2021-05-28
深圳百人科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the above-mentioned target fragment capture methods all require library construction steps before sequencing on the machine. Library construction is often the most important and time-consuming step before sequencing. Its success is directly related to the success of sequencing.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] In an embodiment of the present invention, a two-step PCR technique comprises the following steps:

[0028] The first step is to amplify the specific target: the DNA sample and the specific primer pair mixture are mixed in this part, so that the universal sequences of the forward and reverse primers are respectively connected to the two ends of the specific amplicon determined by the specific primers. On the other hand, this part of the reaction is also called the first step of PCR.

[0029] The second step is to enrich the target: in the enrichment of the target, which is the second step of PCR, we connected the linker after the universal sequence connected in the first step.

[0030] The third step is to mix the library: this part uses the fluorescence quantitative method to quantify the enrichment products, and records the sample information and concentration information.

[0031] The fourth step is to dilute the library: use the fluorescent quantitative method to q...

Embodiment 2

[0037] image 3 It is a graph showing the proportion of data utilization under different methods, where noise represents sequencing noise.

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PUM

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Abstract

The invention discloses a two-step PCR technology, and the technology comprises the following steps: 1, amplifying a specific target spot: mixing a DNA sample and a specific primer pair mixed solution in the part, so universal sequences of forward and reverse primers are respectively connected to two sides of a specific amplicon determined by the specific primers, wherein the part of reaction is also called as first-step PCR; 2, enriching a target spot: connecting a joint behind the universal sequence connected in the first step at the target spot enriching part, namely the PCR part in the second step. And the connected joint is amplified and purified through the second-step PCR, and is mixed into a single test tube for purification. The data utilization rate of the method is higher than that of a sequencing method constructed through a library in the market through simpler operation steps.

Description

technical field [0001] The invention relates to the field of biology, in particular to a two-step PCR technique. Background technique [0002] Polymerase chain reaction, referred to as PCR, is a method for enzymatically synthesizing specific DNA fragments in vitro. It consists of a cycle of high-temperature denaturation, low-temperature annealing (renaturation), and suitable temperature extension. The cycle is carried out so that the target DNA can be rapidly amplified. , has the characteristics of strong specificity, high sensitivity, simple operation, and time saving. It can be used not only for basic research such as gene isolation, cloning and nucleic acid sequence analysis, but also for disease diagnosis or wherever there is DNA or RNA. PCR is also known as cell-free molecular cloning or in vitro primer-directed enzymatic amplification of specific DNA sequences. [0003] DNA probe (DNA probe) is the most commonly used nucleic acid probe. It is single-stranded or doubl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806
CPCC12Q1/6806C12Q2531/113C12Q2525/191C12Q2535/122
Inventor 苗鑫垚沈悦生龚晓娟赵梓丞李梦瑶贺小兰
Owner 深圳百人科技有限公司
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