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Nucleic acid gel electrophoresis testing method

A nucleic acid gel and electrophoresis technology, applied in the field of nucleic acid gel electrophoresis testing, can solve the problems of inability to guarantee the true level of DNA migration, low toxicity, and insufficient sensitivity, etc., and achieve simple and controllable preparation methods, stable molecular structure, and high sensitivity sexual effect

Active Publication Date: 2019-11-08
UELANDY INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

SYBR series dyes have defects such as insufficient sensitivity and low toxicity
The products of American Biotium Co., Ltd. are non-toxic and highly sensitive, but they have a relatively large impact on the migration rate of DNA, that is, the true level of DNA migration cannot be guaranteed, which will lead to deviations in experimental results.
This effect on mobility, the same band displacement will appear to deviate significantly

Method used

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  • Nucleic acid gel electrophoresis testing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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[0032] Put 60.0g 3,8-diamino-6-phenylphenanthridine, 360ml DMF and 37.2ml Pyridine in a 1000ml three-neck round bottom flask, stir mechanically, cool to 0-5°C in an ice-water bath; add 42ml II dropwise, drop Keep warm at 0-5°C during the addition process; continue to react for ten hours until the reaction is completed; filter with suction, add the filter cake to about 2L of pure water, stir mechanically for 30 minutes, and filter with suction; wash the filter cake with 2L of pure water again, Suction filtration to dryness; drying under reduced pressure to constant weight to obtain 48 grams of yellow solid; weigh 347 g of ethyl 6-iodohexanoate, put it in a 2000 ml three-necked flask, add 39.4 g of intermediate I under mechanical stirring, and heat in an oil bath to 100-110°C, react for 3 days until the reaction is complete; cool down to 80°C, add 1500mlEA, reflux for 1h, stop heating, and naturally cool down...

Embodiment 2

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[0041] In a 100 mL three-necked flask equipped with a reflux condenser, add 40 mL of chlorobenzene, and add SM (10 g) into it while stirring. Add dimethyl sulfate (4 mL), mix the three, raise the temperature to reflux state, TLC tracking, developer: methanol (5%) ethyl acetate (95%) and add a small amount of acetic acid, until the end of the reaction (about 2-3 Hour). Add 10 mL of ethanol and continue to reflux for 15 min, stop heating and cool to room temperature, directly filter to remove the solvent, wash the filter cake with ether 2-3 times, and dry the solid in vacuum to obtain the product intermediate I (8 g).

[0042] Add Intermediate I (7 g), 100 mL of acetonitrile, and 20 mL of water to a 250 mL three-necked flask equipped with mechanical stirring and a reflux condenser, gradually heat to reflux, and then reduce iron powder (15 g) and trichloride Iron (200 mg) was added into the reactio...

Embodiment 3

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[0056] Add 2-amino-4,4'-dinitrobiphenyl (17.2 g) and 3-cyanobenzoyl chloride (11 g) into a 250 mL three-necked flask equipped with a reflux condenser, followed by 60 mL of chlorobenzene , stirred by magnetic force and gradually heated to reflux state, and reacted for 4 hours. After the reaction was completed, it was cooled to room temperature, and the chlorobenzene was directly removed by filtration. The obtained filter cake was recrystallized and purified with acetic acid, and the obtained solid was vacuum-dried and dried to obtain the product intermediate I (24.1 g).

[0057] Add Intermediate I (18 g) and 60 mL of nitrobenzene to a 250 mL single-necked flask equipped with a reflux condenser, and add 6 mL of POCl under magnetic stirring 3 , gradually heated to 200°C for 3 hours, then cooled to room temperature, and nitrobenzene and POCl were distilled off under reduced ...

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Abstract

The invention discloses a nucleic acid gel electrophoresis testing method. An adopted dye compound is of a symmetric structure with a bridge connection, and is stable in molecular structure, large incharge carrying amount and large in molecular weight; a preparation method is simple and controllable; when being used as a nucleic acid dye, the dye compound has high sensitivity and almost no influence on the migration rate of DNA, so that the true level of DNA migration can be ensured, and a real test result can be embodied; when the dye compound is used for gel testing, the problem of obviousfluorescence quenching due to the fact that an existing fluorescent dye can form a dimer in an aqueous solution with the high concentration is avoided; and moreover, the compound is large in charge carrying amount and large in molecular weight, so that the compound cannot enter a human body through a cell membrane to cause injury, namely, the compound is nontoxic. Therefore, the nucleic acid gel electrophoresis testing method is highly sensitive, safe and nontoxic.

Description

[0001] The present invention belongs to a divisional application with the title of a compound and its application as a low-mobility nucleic acid dye, the application date is June 13, 2016, and the application number is 2016104106305, and it belongs to the test method part. technical field [0002] The invention belongs to the technical field of biological materials, specifically relates to a compound and its application as a low-mobility nucleic acid dye, and more specifically relates to a nucleic acid gel electrophoresis test method. Background technique [0003] Agarose (nucleic acid) gel electrophoresis is widely used in molecular biology, providing a powerful means for gene recombination, molecular hybridization and DNA sequence research. Fluorescent dyes have been widely used as nucleic acid probes in the past decade. These fluorescent nucleic acid dyes can not only be used in electrophoresis nucleic acid gel staining, but also can be used to quantitatively detect DNA i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447G01N1/30
CPCG01N1/30G01N27/447
Inventor 夏继波焦兆友聂承斌钱近春谈雪良赵育明
Owner UELANDY INC
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