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A method for analyzing nucleic acid sequences by combining samples

A nucleic acid sequence and combined analysis technology, applied in the biological field, can solve the problems of obtaining independent information, unable to infer specific information, etc., and achieve the effect of reducing analysis cost, saving analysis time, and saving time.

Active Publication Date: 2019-03-29
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These 10 sets of independent information cannot be combined to obtain new independent information
Therefore, although it can be known from its "negative" result that the information of all individual samples is "negative", but from its "positive" result, the specific information of the information of a single sample cannot be inferred

Method used

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  • A method for analyzing nucleic acid sequences by combining samples
  • A method for analyzing nucleic acid sequences by combining samples
  • A method for analyzing nucleic acid sequences by combining samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: DNA chip typing based on sample combination mitochondrial mutation site C3206T site

[0042] 1. Genomic DNA sample preparation

[0043] First, number 100 human blood samples, numbered from 1, 2, 3, ..., 100, group samples numbered 1-45 into 10 groups according to Table 1-1, and group numbers 46-100 according to Table 1-2 Divide into 11 groups, a total of 21 groups and mark the corresponding group number. Then the 21 groups of mixed blood samples were extracted with mitochondrial DNA extraction kit (mtDNAExtractor (R) WB KIT, WAKO), and the following steps 2-7 were performed on the 21 groups of mixed samples according to the following steps.

[0044] 2. PCR

[0045] 50 μL PCR amplification system contains: 200 ng genomic DNA, 0.2mM dNTP, 0.8 μM acrylamide-modified forward primer 5'-GCAGCCGCTATTAAAGGTTCG and 0.8 μM reverse primer 5'-GGGCTCT GCCATCTTAACAAA, 2 U Taq DNA polymerase, 1X Amplification Buffer, 1.5 mM MgCl2. Amplification conditions: pre-denaturat...

Embodiment 2

[0058] Example 2: Pyrosequencing typing of mitochondrial mutation site A5301G site based on sample combination

[0059] 1. Genomic DNA sample preparation

[0060] First, 56 human blood samples are numbered from 1, 2, 3, ..., 100, and then combined and grouped according to Table 2, and marked as , , ,..., , divided into 8 groups. Then the 8 groups of mixed blood samples were extracted with mitochondrial DNA extraction kit (mtDNA Extractor (R) WB KIT, WAKO), and the following steps 2 to 5 were performed on the 8 groups of mixed samples according to the following steps.

[0061] 2. PCR

[0062] 50 μL PCR amplification system contains: 200 ng mitochondrial DNA, 0.2mM dNTP, 0.8 μM forward primer 5'-biotin-CAATTACCCCATAGGATGA and 0.8 μM reverse primer 5'-AGGCGTAGGTAGAAG TAGAGGTT, 2 UTaq DNA polymerase, 1× amplification Buffer, 1.5 mM MgCl 2 . Amplification conditions: pre-denaturation at 94°C for 5 minutes, 35 thermal cycles (denaturation at 94°C for 30 seconds-annealing ...

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Abstract

The invention provides a method for combinatory analysis of nucleic acid sequence of a specimen. The method comprises the following steps: dividing a plurality of specimens to be analyzed into a plurality of groups according to a specific combination mode, ensuring that each specimen is analyzed for y times in different y (y is greater than or equal to 2) groups and only in y groups, analyzing the nucleic acid sequence of each group serving as one analysis sample, and finally according to the analysis results of the plurality groups of samples, judging the analysis result of a specific specimen in a combination mode. The method aims at a characteristic nucleic acid fragment sequence of a single index, moreover only a few specimens of multiple specimens have the characteristic nucleic acid fragment sequence, and the purpose of the method is to confirm specimens with the characteristic nucleic acid fragment sequence by analyzing samples of which the number is far smaller than that of the specimens in the combination mode. By adopting the method, the analysis efficiency of the specimens can be greatly improved, and the analysis cost can be greatly reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for analyzing specific nucleic acid sequence fragments by greatly reducing the number of samples to be analyzed through a combined method. Background technique [0002] The development and completion of the Human Genome Project and various model organism genome projects has brought mankind into the post-gene era, which has had a huge impact on contemporary biological and medical research, and the related disciplines of molecular biology have developed rapidly. It will become possible to understand the differences of life, the law of disease occurrence and development, and the interaction between drugs and living organisms at the genetic level. As far as gene sequence analysis is concerned, the focus of the post-genome era has shifted from whole-genome sequence determination to the comparison of individual genetic differences and inter-species genetic differences ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G16B30/00
CPCG16B30/00
Inventor 肖鹏峰孙啸
Owner SOUTHEAST UNIV