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A Method for Identifying All Chromosomes of Poplar Using Oligonucleotide Probes

An oligonucleotide probe and chromosome technology, which is applied in the field of identifying all the chromosomes of poplar by using oligonucleotide probes, can solve the problems of lagging cytogenetic research, difficulty in accurately identifying chromosomes, etc., and achieve good experimental repeatability. , high-resolution effect

Active Publication Date: 2019-12-20
NANJING FORESTRY UNIV
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Problems solved by technology

However, what does not match the status of the poplar model tree species is that there is no high-resolution karyotype map for research at home and abroad
This is mainly because the poplar chromosomes are small, and it is difficult to accurately identify each chromosome based on the shape of the mitotic metaphase chromosomes, resulting in a significant lag in cytogenetic research

Method used

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  • A Method for Identifying All Chromosomes of Poplar Using Oligonucleotide Probes

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Embodiment 1

[0036] A kind of method utilizing oligonucleotide probe to identify poplar whole chromosome, comprises the steps:

[0037] (1) Chromosome production

[0038] Collect young poplar root tips of 1 cm, immerse in 2mM 8-hydroxyquinoline solution for pretreatment at room temperature in the dark for 2.5 hours; rinse with water for 10 minutes, transfer to freshly prepared Carnot’s fixative, and store at -20°C until use ;

[0039] The root tip taken out of the fixative was rinsed in water for 10 minutes, the meristem part was cut with a double-sided blade, and the root tip meristem part was enzymatically hydrolyzed in an enzyme mixture of 4% cellulase and 2% pectinase at 37°C 1h; put the root tip meristem into deionized water to terminate the reaction;

[0040] Use a pipette to place the root tip on a clean glass slide, absorb excess water with a filter paper strip; mash the root tip tissue with tweezers and a dissecting needle; quickly add 20 μl of 60% glacial acetic acid dropwise, ...

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Abstract

The invention discloses a method for identifying all the chromosomes of poplar by using oligonucleotide probes. According to the genome information of Populus trichocarpa, the specific oligonucleotide sequence of each chromosome is obtained through bioinformatics analysis, and the synthetic poplar 1 The oligonucleotide library corresponding to chromosome 19 is then labeled with biotin or digoxin into oligonucleotide probes, and then the chromosomes of poplar are identified by fluorescence in situ hybridization technology. Each round of hybridization can be used at most at the same time The oligonucleotide probes of the three chromosomes were repeated 7 times on the same metaphase division phase, and the 19 pairs of chromosomes of Populus species could be accurately identified. It has the advantages of common use in Populus species, good experimental repeatability, and high resolution; the invention provides a new method for accurately identifying each chromosome of Populus species, and has laid a solid foundation for the molecular cytogenetics research of Populus Base.

Description

technical field [0001] The invention relates to the field of molecular cytogenetics, in particular to a method for using oligonucleotide probes to identify all the chromosomes of poplar trees. Background technique [0002] Fluorescence in situ hybridization (FISH) is a molecular cytogenetic technique developed in the 1980s. FISH technology can directly locate the DNA sequence on the chromosome, which provides an efficient method for the construction of cytological maps. This technology only needs to use the specific DNA sequence of each chromosome as a probe, and can visually distinguish each chromosome. At present, FISH technology has been widely used in the karyotype analysis of various plants. Chromosome painting is to prepare a probe from the DNA of the whole chromosome, a certain chromosome arm (long arm or short arm) or a certain fragment of the chromosome, and use the FISH method to hybridize the probe to the chromosome. It is a new technology to analyze and study ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6841
Inventor 席梦利辛昊阳赵乙琏尚大鑫宁仪杭
Owner NANJING FORESTRY UNIV
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