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A kind of oligonucleotide dye solution and its application and usage

An oligonucleotide and oligonucleotide probe technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem that dye solution cannot comprehensively identify species chromosomes, etc., and achieve fast detection time and cost. low effect

Active Publication Date: 2019-10-01
JIANGSU ACAD OF AGRI SCI +1
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Problems solved by technology

[0004] The purpose of the present invention is to solve the problem that the dye solution composed of the existing single probe cannot fully identify the species chromosome, and provide an oligonucleotide dye solution capable of fully identifying the species chromosome and its usage and application

Method used

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  • A kind of oligonucleotide dye solution and its application and usage
  • A kind of oligonucleotide dye solution and its application and usage
  • A kind of oligonucleotide dye solution and its application and usage

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Embodiment

[0025] 1. Plant material and chromosome preparation

[0026] A total of 16 maize materials were used for chromosome analysis, including 3 inbred lines and 2 corresponding hybrids, and other 11 main hybrids in China (Table 1). Corn seeds were soaked in water and cultured overnight at 28°C until the root tip length reached 2 cm. Cut the root tip into a 2ml centrifuge tube, and treat it with 0.29g / L 8-hydroxyquinoline solution for 3-4 hours. Then use ethanol glacial acetic acid solution with a volume ratio of 3:1 to fix for 2-3 days. When preparing chromosomes, cut out 0.3-0.5 mm of root tip tissue and add 45% (v / v) glacial acetic acid solution for dripping, and freeze overnight at -70°C. Slides containing mitotic chromosomes were dehydrated with 100% ethanol for FISH or ND-FISH analysis.

[0027] Table 1 Materials used

[0028]

[0029]

[0030] 2. Development and labeling of single-stranded oligonucleotide (SSON) probes

[0031] SSON probes were developed using simpl...

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Abstract

The invention belongs to the field of cytobiology and molecular biology studies and particularly relates to oligonucleotide dye liquor and application and a usage method thereof. The oligonucleotide dye liquor comprises YMPNS-Centc69-1, mknob-2, YMPNS-MR68-3 and (ACT)<10> which are four modified oligonucleotide probes, mixing is conducted according to a certain ratio, dilution is conducted through 2xSSC, and thus the oligonucleotide dye liquor is formed. 8 [mu]l of the oligonucleotide dye liquor is added onto each non-denatured chromosome piece, 2.5 [mu]l of 50%w / v dextran sulfate is added, hybridization is conducted in an incubator at the temperature of 37 DEG C for two hours, then washing is conducted in 2xSSC at the temperature of 42 DEG C for five minutes, and observation can be conducted below a fluorescence microscope after over dyeing through DAPI. The oligonucleotide dye liquor can be developed into a commodity to be used for fast chromosome identification of corn and affinity species of the corn.

Description

technical field [0001] The invention belongs to the research fields of cell biology and molecular biology, and in particular relates to an oligonucleotide dye solution and its application and usage. Background technique [0002] Oligonucleotides are short DNA or RNA sequences that can be synthesized artificially. Oligonucleotides are widely used as probes for chromosome identification. Due to the high sensitivity and resolution of single-strand oligonucleotide (SSON) probes, they have been applied in the chromosome identification of wheat, rye and other crops as well as humans (Fu et al. 2015, Du et al. 2016, Beliveau et al. 2012). The characteristic of SSON probe is that it is easy to synthesize and label the detection signal, which can avoid the denaturation step in the conventional fluorescence in situ hybridization (fluorescence in situ hybridization, FISH) process. This method is generally called non-denaturing fluorescence in situ hybridization (non-denaturing FISH,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6841
CPCC12Q1/6841C12Q2563/107C12Q2543/10
Inventor 张体付赵涵亓增军朱敏秋杜培庄丽芳
Owner JIANGSU ACAD OF AGRI SCI
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