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Kit for simultaneously extracting DNA and RNA and using method thereof

A kit and reagent technology, applied in the field of genetic engineering, can solve problems such as complex operation and difficulty in ensuring specific elution of RNA

Pending Publication Date: 2020-10-23
广州捷倍斯生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The invention is complicated to operate and uses toxic organic solvents such as phenol chloroform, and the method is to elute the RNA from the total nucleic acid, because the RNA is more closely adsorbed to the silicon material, so it is difficult to ensure the specific elution of the RNA. Does not elute DNA

Method used

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  • Kit for simultaneously extracting DNA and RNA and using method thereof
  • Kit for simultaneously extracting DNA and RNA and using method thereof
  • Kit for simultaneously extracting DNA and RNA and using method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0040] This embodiment provides a kit for simultaneously extracting DNA and RNA, comprising the following components:

[0041] Cell lysate: 2% SDS, 100mM Tris, 2% PVP, 0.5% mercaptoethanol, 1.4M NaCl, 10mM EDTA;

[0042] Nucleic acid binding solution: 75% ethanol solution;

[0043] Nucleic acid adsorption material: silica purification column;

[0044] Eluent A: 0.1M sodium chloride, 30% ethanol;

[0045] Protein-removing rinse solution: 1M guanidine hydrochloride 50% ethanol;

[0046] Desalting rinse solution: 75% ethanol solution;

[0047] Eluent B: DEPC treated water.

[0048] The method for simultaneously extracting DNA and RNA kit described in this implementation comprises the following steps:

[0049] (1) Add 100mg of plant samples to a 1.5ml centrifuge tube containing 0.6ml of cell lysate after grinding with liquid nitrogen, vigorously vortex and mix well, after 10 minutes in a water bath at 70°C, add 0.2ml of 3M ammonium acetate solution, mix Centrifuge at 12000rp...

Embodiment 2

[0058] A kit for simultaneously extracting DNA and RNA, comprising the following components:

[0059] Cell lysate: 4M guanidine isothiocyanate, 100mM Tris, 30mM TCEP, 1% Triton X-100, 10mM EDTA;

[0060] Nucleic acid binding solution: 75% ethanol solution;

[0061] Nucleic acid adsorption material: silica purification column;

[0062] Eluent A: 0.1M ammonium chloride, 40% ethanol;

[0063] Protein-removing rinse solution: 3M guanidine isothiocyanate, 40% ethanol;

[0064] Desalting rinse solution: 75% ethanol solution;

[0065] Eluent B: DEPC treated water.

[0066] The method for using the above-mentioned simultaneous extraction of DNA and RNA kit comprises the following steps:

[0067] (1) Add 30 mg of animal tissue samples to a 1.5 ml centrifuge tube containing 0.6 ml of cell lysate, grind the tissue thoroughly with a grinding pestle, mix well and centrifuge at 12,000 rpm for 1 min at room temperature to obtain the supernatant;

[0068] (2) Take 0.4 ml of the supernat...

Embodiment 3

[0075] A kit for simultaneously extracting DNA and RNA, comprising the following components:

[0076] Cell lysate: 5M guanidine hydrochloride, 0.1M Tris, 1% Triton X-100, 4mg / ml proteinase K;

[0077] Nucleic acid binding solution: 3M guanidine isothiocyanate, 60% ethanol;

[0078] Nucleic acid adsorption material: siliceous purification column and silica ferric oxide nano-magnetic beads;

[0079] Eluent A: 1M sodium chloride, 0.1M ammonium chloride, 35% ethanol;

[0080] Protein-removing rinse solution: 3M guanidine isothiocyanate, 40% ethanol;

[0081] Desalting rinse solution: 80% ethanol solution;

[0082] Eluent B: DEPC treated water.

[0083] The method for using the above-mentioned simultaneous extraction of DNA and RNA kit comprises the following steps:

[0084] (1) Take 1ml of microbial culture solution and place it in a 1.5ml centrifuge tube, centrifuge at 8000rpm for 1 minute at room temperature to collect the bacteria, and remove the culture medium;

[0085] ...

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Abstract

The invention relates to a kit for simultaneously extracting DNA and RNA and a using method thereof. The kit comprises the following reagents: a cell lysis solution, a nucleic acid binding solution, anucleic acid adsorption material, an eluate A, a deproteinized rinsing solution, a demineralized rinsing solution and an eluate B. According to the method, the DNA is selectively eluted, and the RNAis continuously retained on the adsorption material, so that the effect of fractionating the DNA from the RNA is achieved. Different from a TRIzol method for simultaneously extracting DNA and RNA, themethod provided by the invention uses few reagents, does not use phenol, chloroform and other toxic solvents, can be used on a nucleic acid purification instrument, and has the advantages of rapidness, high efficiency, high flux, environmental protection and capability of automatically and simultaneously extracting DNA and RNA.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a kit for simultaneously extracting DNA and RNA and a use method thereof. Background technique [0002] DNA and RNA are collectively referred to as nucleic acid. Nucleic acid is the most basic research object in the research and application of molecular biology. How to obtain high-quality nucleic acid has become an important and basic experimental technique. The current technical routes are all aimed at extracting DNA or RNA separately. If you want to extract DNA and RNA at the same time, you must divide the experimental materials into two. The technologies currently used for nucleic acid extraction mainly include: phenol-chloroform extraction, silica gel column separation, salting-out and magnetic bead methods. Basically, these techniques can only extract DNA or RNA, and cannot use the same sample to extract DNA and RNA at the same time. [0003] The classic method for extra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/101C12N15/1013
Inventor 卢秀劲王小芳
Owner 广州捷倍斯生物科技有限公司
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