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A method for the detection of loquat aneuploid molecular karyotype based on ssr markers and qPCR

A technology of aneuploidy and molecular nucleus, which is applied in the field of loquat aneuploid molecular karyotype detection based on SSR markers and qPCR, can solve the problems of complexity, error-prone, and high SNP cost, and achieve low cost, short time, Wide range of effects

Active Publication Date: 2020-02-14
SOUTHWEST UNIV
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  • Summary
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The first two methods rely on more complex chromosome preparation and chromosome morphology observation, which are prone to errors; QF-PCR and HRM detection are limited to heterozygous genotypes; MLPA and CGH are more complicated in probe preparation; high-density SNP costs are higher

Method used

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  • A method for the detection of loquat aneuploid molecular karyotype based on ssr markers and qPCR
  • A method for the detection of loquat aneuploid molecular karyotype based on ssr markers and qPCR
  • A method for the detection of loquat aneuploid molecular karyotype based on ssr markers and qPCR

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Experimental program
Comparison scheme
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Embodiment 1

[0026] 1. Experimental steps:

[0027] (1) Take the tested materials ('A322-1', 'A322-2', 'A322' of 'A322-1', 'A322-2', 'A322' of 'A313-1' of 'H39', 'A313' of 'Nuclear National Jade' -3') and the young leaves of the control material (comprising the male parent and the female parent of the tested material);

[0028] (2) Extract loquat genomic DNA, use 1% agarose gel to detect DNA quality, and measure the concentration and then dilute to 50ng / μL;

[0029] (3) qPCR detection of loquat genomic DNA in (3) was performed using 17 pairs of SSR primers (Table 1) obtained by screening.

[0030] 2. Data processing

[0031] (1) Amplification curve (eg, figure 1 shown) select 3 CT values ​​during the exponential growth phase of PCR amplification (bottom (CT 1 ), Central (CT 2 ), top (CT 3 ));

[0032] (2) View the amount of PCR product of each pair of SSR primers at the three CT values ​​(ΔRn 1 , ΔRn 2 , ΔRn 3 ), and calculate the average value of ΔRn for each pair of SSR primer...

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Abstract

The invention provides a method of detecting aneuploidy molecular karyotype of loquat based on SSR (simple sequence repeat) markers and qPCR (quantitative polymerase chain reaction). Seventeen SSR primers acquired by screening are suitable for detecting complete molecular karyotype of aneuploidy of loquat in combination with the qPCR technique. extra reference genes are not required herein; the method is applicable to both hybrid and homozygous genotypes, is not limited to material and chromosome morphology, is low in cost and time consumption and is widely applicable.

Description

technical field [0001] The invention relates to the identification of loquat chromosome karyotype, in particular to a method for detecting loquat aneuploid molecular karyotype based on SSR markers and qPCR. Background technique [0002] Loquat (Eriobotrya japonica Lindl) is a plant of the genus Loquat in the subfamily Rosaceae (Rosaceae), and is widely planted in subtropical regions of my country and other Asian countries. Loquat originated in my country, has a cultivation history of more than 2,000 years, and is rich in germplasm resources. Loquat is a subtropical evergreen fruit tree. The mature loquat fruit is mostly spherical or oval, soft and juicy, and rich in glutamic acid, phenolic substances (especially flavonoids), vitamins and carotenoids. health fruit. In addition to eating fresh, loquat fruit is also a good raw material for processing canned food, jam, fruit paste, jelly and fruit wine. [0003] Aneuploid (aneuploid) refers to a biological individual with one...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 温国党江波郭启高梁国鲁何桥蒋朋飞
Owner SOUTHWEST UNIV