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Molecular weight internal lane standard and application thereof
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A molecular weight internal standard, molecular weight technology, applied in the new molecular weight internal standard and its application, molecular weight internal standard field, to ensure the correct typing and easy interpretation.
Inactive Publication Date: 2016-02-03
SUZHOU NUHIGH BIOTECH
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[0004] The present invention aims at the deficiencies existing in the use of the existing molecular weight internal standard, and provides a molecular weight internal standard and its application that can clearly distinguish a single base and ensure correct typing, thereby facilitating the interpretation of operators
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Embodiment 1
[0028] Embodiment 1: the preparation method of novel molecular weight internal standard described in the present invention
[0029] The molecular weight internal standard of the present invention can be prepared by polymerasechain reaction (PCR) or enzymedigestion method, and the following two methods are described in detail respectively, so that those skilled in the art can implement without creative work.
[0031] 1.1 Design and synthesis of upstream and downstream primers
[0032] via a widely used exogenous DNA template. Use PrimerExpress software to analyze the complete sequence of exogenous DNA, design primers upstream (5'), and mark with NH635 at the 5' of the primers. Upstream primer 5'-GCCCGTCGAGAATACTGGCAATTTCACCTGC-3', annealing temperature 58°C. Take the upstream primer as the starting point, and find the end point of the downstream primer at the corresponding distance downstream. The length and sequence of the downstream...
[0047] The molecular weight internal standard prepared according to the method described in Example 1 has a molecular weight of 75, 76, 80, 100, 140, 160, 161, 175, 180, 200, 225, 226, 250, 275, 300, 330, 331 , 360, 390, 445, 500, 501bp fragments.
[0048] When a certain part of the Buffer in the Buffer tank, the water in the water tank, the capillary, and the POP glue and formamide has expired or failed, use the molecular weight internal standard of the present invention to carry out STR typing detection, and the results are as follows figure 1 shown.
[0049] figure 1 The results show that the molecular weight internal standard of the present invention can distinguish the single base difference between 160bp and 161bp, 225bp and 226bp, ensuring single base resolution; while the LIZ-500 molecular weight internal standard cannot distinguish between 160bp and 161bp, nor can it separate 225bp and 226bp , cannot resolve single bases,...
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Abstract
The invention belongs to the technical field of biology and discloses a molecular weight internal lane standard. The molecular weight internal lane standard comprises n nucleotide fragments with recognizable markers and different molecular weights, wherein the n nucleotide fragments include at least p nucleotide fragment pairs differing by one basic group respectively, in other words, the basic group number of one fragment of any nucleotide fragment pair differing by one basic group is m, and the basic group number of the other fragment is m+1 or m-1. Compared with existing commercial molecular weight internal lane standards, the molecular weight internal lane standard has the advantages that the molecular weight internal lane standard is capable of distinguishing single basic group, effectively differentiating PCR (polymerasechain reaction) product fragments differing by one basic group and guaranteeing correct typing to facilitate interpretation of detection personnel, thereby being widely applicable to STR typing, sequencing, large-fragment analysis and genome analysis.
Description
technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a novel molecular weight internal standard and its application. In particular, it relates to a molecular weight internal standard capable of clearly distinguishing single bases and ensuring correct typing and its application Background technique [0002] Genome scanning is a method of finding markers closely linked to specific traits or genes from the genetic markers of the whole genome, and locating the related genes on the chromosome. Genotyping (Genotyping), also known as Genotype Analysis (Genotypicassay), is a technology that uses biological detection methods to determine an individual's genotype (Genotype), which uses technologies including polymerasechain reaction (PCR), DNA fragment analysis, Oligonucleotide probes (ASOprobes), gene sequencing, nucleic acid hybridization, genechip technology, etc. Gene scanning and genotyping technologies have developed rapidl...
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