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Method and equipment for detecting cytoplasmic inheritance

A cytoplasmic and sample technology, applied in the field of cytoplasmic genetic detection, can solve problems such as misjudgment and achieve the effect of flexible sampling

Active Publication Date: 2018-12-28
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the multi-copy nature of cytoplasmic DNA, traditional markers can only detect cytoplasmic DNA from a single parent due to differences in template concentration and PCR competition (Wang et al., 2010), resulting in misjudgment

Method used

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  • Method and equipment for detecting cytoplasmic inheritance
  • Method and equipment for detecting cytoplasmic inheritance
  • Method and equipment for detecting cytoplasmic inheritance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Arabidopsis thaliana mitochondrial DNA genetic mode detection, including the following steps:

[0063] (1) According to literature reports (Matsushima et al., 2011), Arabidopsis ecotypes C24 and Col have a SNP site in the genome sequence of the mitochondrial gene matR. Therefore, for this SNP site, use Primer Express3.1 software, based on TaqMan MGB Allelic Discrimination mode, design primers and probes for paternal and maternal materials.

[0064] Wherein, the obtained mitochondrial genome sequences of C24 and Col are respectively shown as SEQ ID NO: 1 and SEQ ID NO: 2, and there is a SNP (SNP.C29A) at 29 bp compared with the two.

[0065] The front and back primer sequences of the two parents designed for this sequence are consistent, as shown in SEQ ID NO: 3 and SEQ ID NO: 4, the sizes are 26 and 20 bp respectively, and the size of the amplified fragment is 68 bp.

[0066] The specific probe sequence designed for C24 is shown in SEQ ID NO:5, the first one at the 5' ...

Embodiment 2

[0076] The detection of the mode of inheritance of cucumber mitochondrial DNA comprises the following steps:

[0077] (1) Comparatively analyze the mitochondrial genome polymorphisms of the parent materials '9930' and 'B', find suitable polymorphic sites, and then design synthetic primers and probes for the polymorphic sites, and the probes span the polymorphisms sites and labeled with FAM and HEX fluorophores, respectively. Among them, the parental materials 9930 and B are inbred line materials kept in the laboratory, and their mitochondrial genome sequences were respectively obtained by next-generation sequencing, and compared with the published reference mitochondrial genome of cucumber 'Calypso' by software Bowtie 2, bam files were generated , and load the bam file into IGV 2.4.5, and compare the sequencing results to find effective polymorphic sites. Then, using Primer Express 3.1 software, based on the TaqManAllelic Discrimination model, primers and probes for the pater...

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Abstract

The invention relates to the technical field of biology and in particular relates to a method and equipment for detecting cytoplasmic inheritance. The method for detecting cytoplasmic inheritance provided by the invention comprises the following steps: (1) respectively obtaining different polymorphic sites in parental samples of biological materials based on polymorphism of cytoplasmic nucleic acid sequences of the parental samples of biological materials, wherein the parental samples comprise male parent samples and female parent samples; (2) obtaining polymorphism results of cytoplasmic nucleic acid sequences of offspring samples by utilizing a digital PCR (Polymerase Chain Reaction) technology based on different polymorphic sites; (3) determining the cytoplasmic inheritance of the biological materials based on the polymorphism results in cytoplasm of the offspring samples. Meanwhile, the invention correspondingly provides a device for detecting cytoplasmic inheritance. The method and device provided by the invention are used for detecting the cytoplasmic inheritance characteristics, are flexible in sampling, and have high resolution, and the competitive effect existing between templates is avoided; meanwhile, the method and device have extremely high sensitivity.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method and equipment for detecting cytoplasmic inheritance. Background technique [0002] Cytoplasmic inheritance refers to the genetic phenomenon determined by genetic factors outside the nucleus, which is called non-Mendelian inheritance or extranuclear inheritance. Cytoplasmic inheritance in animals is mainly mitochondrial inheritance, while cytoplasmic inheritance in plants includes mitochondrial inheritance and plastid inheritance. In plants, the earliest non-Mendelian inheritance phenomenon was reported in 1909. When Baur et al. studied the inheritance of "leaf spots" in geraniums, they found that the inheritance of the offspring's parental leaf spots did not show the Mendelian inheritance method (Baur, 1909 ). The mode of cytoplasmic inheritance is closely related to plant cytoplasmic male sterility, growth retardation caused by plastid or mitochondrial genome mutations, e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6858C12Q1/6895C12M1/34C12M1/00
CPCC12Q1/6858C12Q1/6895C12Q2600/156C12Q2531/113C12Q2563/107
Inventor 沈佳张跃建寿伟松
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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