Preparation method, amplification primer and detection reagent of hpv full-length genome quality control product

A technique for amplifying primers and genomes, which is applied in biochemical equipment and methods, microbiological determination/inspection, recombinant DNA technology, etc. Higher price and other issues, to achieve good applicability, avoid low connection efficiency, and controllable concentration

A technique for amplifying primers and genomes, which is applied in biochemical equipment and methods, microbiological determination/inspection, recombinant DNA technology, etc. Higher price and other issues, to achieve good applicability, avoid low connection efficiency, and controllable concentration

CN108929869BActive Publication Date: 2022-03-08亚能生物技术(深圳)有限公司
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  • Preparation method, amplification primer and detection reagent of hpv full-length genome quality control product
  • Preparation method, amplification primer and detection reagent of hpv full-length genome quality control product
  • Preparation method, amplification primer and detection reagent of hpv full-length genome quality control product

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Experimental program
Comparison scheme
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Embodiment

[0110] see Figure 1-8 , Figure 1-2 Electrophoresis results for electrophoresis of PCR products after sample DNA amplification; image 3 It is a schematic diagram of the structure of TVector (including the cloning site); Figure 4 Select the fluorescence profile for the sample, Figure 4 The middle curve corresponds to standard 1, sample, standard 2, standard 3 and standard 4 from left to right; Figure 5 is the standard curve formula graph; Image 6 The initial Ct value map for the stability study; Figure 7 Ct value change chart for stability research; Figure 8 It is the chart of the value determination results of 23 quality control substances by digital PCR absolute quantification method.

[0111] The preparation method of the human papillomavirus full-length genome quality control product of the present embodiment comprises:

[0112] (1) HPV nucleic acid extraction and type determination

[0113] Use the nucleic acid extraction reagent of Yaneng Biotechnology (Sh...

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Abstract

The invention relates to a preparation method of a HPV full-length genome quality control product, an amplification primer and a detection reagent. Wherein, the preparation method of the HPV full-length genome quality control product comprises the following steps: (1) obtaining a DNA template; (2) using amplification primers to perform PCR amplification; (3) using HD Cloning connection technology to connect the target DNA fragment to the carrier ; (4) introducing the plasmid into competent cells, and then extracting the plasmid; (5) diluting the plasmid with human genome. The preparation method of the human papillomavirus full-length genome quality control product of the present invention can successfully obtain the HPV gene detection quality control product that has no biological infection risk type, can simulate clinical samples, can be produced in large quantities, has a controllable concentration, and is stably stored. Has good applicability.

Description

technical field [0001] The invention relates to gene detection technology, in particular to a preparation method of HPV full-length genome quality control product, amplification primers and detection reagents. Background technique [0002] Human papillomavirus (Human Papillomavirus, HPV) is a group of non-enveloped small DNA viruses, belonging to Papillomaviridae. The morphological feature of the HPV virus is usually an icosahedron with a diameter of 52-55 nm. The length of its genomic DNA is about 8000bp, and the genome is functionally divided into three parts: including early transcription regions (coding early proteins such as E1, E2, E4, E5, E6, E7, etc., involved in virus replication, transcription, translation regulation and transformation, etc. function), late transcribed region (encodes major capsid protein L1 and minor capsid protein L2 for virus assembly; L1 accounts for about 80% of the capsid protein and is highly conserved, L2 has less content and more variatio...

Claims

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Application Information

Patent Timeline
08 Mar 2022
Publication
CN108929869B
IPC
C12N15/11; C12Q1/6806; C12Q1/70; C12Q1/6851; C12N15/70
CPC
C12N15/11; C12N15/70; C12Q1/6806; C12Q1/6851; C12Q1/708; C12Q2531/113; C12Q2563/107
Inventors
卢舟宇; 田洁