Kit used for RNA extraction and extraction method thereof

An extraction method and kit technology, applied in DNA preparation, recombinant DNA technology and other directions, can solve the problems of poor RNA integrity, expensive kits, complicated operations, etc., and achieve easy and widespread application, low cost, and simple extraction steps. Effect

Inactive Publication Date: 2020-03-31
北京华颉基因医疗技术有限公司
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Problems solved by technology

Existing RNA extraction methods have complex operations, low extraction rates, and poor RNA integrity, or the ex

Method used

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  • Kit used for RNA extraction and extraction method thereof
  • Kit used for RNA extraction and extraction method thereof
  • Kit used for RNA extraction and extraction method thereof

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Embodiment Construction

[0023] The technical solution adopted in this specific embodiment is: an RNA extraction kit includes the following reagent components: diethyl pyrophosphate, sterile double distilled water, guanidine isothiocyanate, sodium dodecylsulfonate, polyethylene Pyrrolidone, β-mercaptoethanol, NaAC, water-saturated phenol, chloroform-isoamyl alcohol, ethanol, glycerol, and sorbitol. The pH value in the extraction kit is between 5.0-5.3, most preferably 5.2.

[0024] Described guanidine isothiocyanate, NaAC, glycerin, sorbitol concentration are respectively 3-4.5M, 1.5-2.5M, 0.001-0.010% V / V, 0.8-1.5M / L, and the pH value of NaAC is at 3.5- Between 4.5, ethanol was prepared with 75% absolute ethanol, diethyl pyrophosphate was used to prepare RNase-free double-distilled water, and RNase-free double-distilled water was used to prepare reagents.

[0025] An extraction method of an RNA extraction kit comprises the following steps: Step 1, taking a certain amount of tissue, adding polyvinylp...

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Abstract

The invention provides a kit used for RNA extraction and an extraction method thereof. The kit used for RNA extraction comprises diethyl pyrocarbonate, sterile double distilled water, guanidinium isothiocyanate, sodium dodecyl sulfate, polyvinylpyrrolidone, beta-mercaptoethanol, NaAC, phenol water, chloroform-isoamylol, ethyl alcohol, glycerinum and sorbitol. By means of the technical scheme, thekit has the advantages that the design is reasonable, tissue RNA of the human bodies, mice and tobaccos can be well extracted, the extraction steps are simple, the cost is low, and the kit is easy towide apply. The required RNA sample size is low, the RNA extraction rate is high, the integrity of RNA obtained through extraction is very high, and the kit is completely suitable for downstream research work, such as sequencing, RT-PCR and N-count.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for RNA extraction and an extraction method thereof. Background technique [0002] With the rapid development of molecular biology, there are more and more studies on nucleic acids, and tissue samples are widely used in medical research, especially in clinical diagnosis. Extracting high-quality total RNA from tissue samples is the basis for downstream sequencing, qPCR, chip detection and other technologies, but there are technical difficulties in extracting high-quality total RNA. First, RNA extraction should prevent RNase contamination. RNase is ubiquitous. In any step of the experimental operation, any accidental negligence or improper operation may cause RNase contamination, which will lead to the failure of the entire experiment (Shaban N M, Harvey S, Perrino F W, et al. The Structure of the Mammalian RNase H2Complex Provides Insight into RNA·DNA Hybrid Processing to Prev...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 葛晓颉彦华
Owner 北京华颉基因医疗技术有限公司
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