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Ribonucleic acid (RNA) extraction kit without deoxyribonucleic acid (DNA) residues and RNA extraction method

An extraction method and kit technology, applied in DNA preparation, recombinant DNA technology and other directions, can solve problems such as DNA residues, achieve the effects of less residual DNA, simplify steps and operation time, and solve genomic DNA residues

Active Publication Date: 2012-10-03
BEIJING AIDLAB BIOTECH
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  • Claims
  • Application Information

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Problems solved by technology

[0006] In view of the deficiencies in the prior art, the object of the present invention is to provide a RNA extraction kit and RNA extraction method without DNA residues without using toxic substances such as phenol and chloroform, and can effectively solve the problem of DNA residues in RNA extraction and purification. question

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  • Ribonucleic acid (RNA) extraction kit without deoxyribonucleic acid (DNA) residues and RNA extraction method
  • Ribonucleic acid (RNA) extraction kit without deoxyribonucleic acid (DNA) residues and RNA extraction method

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Embodiment 1

[0035] Embodiment 1. extract the RNA of lily petal, pistil, stamen

[0036] (1) Take 50 mg of lily flower petals, 50 mg of lily flower pistils, and 50 mg of lily flower stamens, and homogenize them with 0.5 mL of lysate to obtain a lysate. The lysate contains Tris-HCl with a concentration of 10 mmol / L and a pH of 7.0, and guanidine isothiocyanate with a concentration of 3 mol / L. The preparation method of the lysate is: weigh 35.45 grams of guanidine isothiocyanate, add an appropriate amount of ribonuclease-free water to dissolve, then add 1 mL of Tris-HCl with a pH of 7.0 and a concentration of 1 mol / L, mix well, and set the volume to 100 mL .

[0037] (2) Transfer the lysate into a centrifuge tube, shake vigorously for 15 seconds, centrifuge at 13000rpm for 5-10 minutes to take the supernatant, add 1 / 2 volume of absolute ethanol of the supernatant and mix well, transfer to a DNA adsorption column for further analysis Adsorption centrifugation. The DNA adsorption column is ...

Embodiment 2

[0043] Embodiment 2. extract the RNA of grass stem

[0044] (1) Homogenize 50 mg of herbaceous stems with 0.5 mL of lysate to obtain a lysate. The lysate contains Tris-HCl with a concentration of 10 mmol / L and a pH of 8.0, and guanidine isothiocyanate with a concentration of 4 mol / L. The preparation method of the lysate is: weigh 47.26 grams of guanidine isothiocyanate, add an appropriate amount of ribonuclease-free water to dissolve, then add 1 mL of Tris-HCl with a pH of 8.0 and a concentration of 1 mol / L, mix well, and dilute to 100 mL.

[0045] (2) Transfer the lysate into a centrifuge tube, shake vigorously for 15 seconds, centrifuge at 13000rpm for 5-10 minutes to take the supernatant, add 1 / 2 volume of absolute ethanol to the supernatant, mix well and transfer to a DNA adsorption column for further analysis Adsorption centrifugation. The DNA adsorption column is a silicon membrane adsorption column.

[0046] (3) Add 0.5mL RNA eluent to the DNA adsorption column, cent...

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Abstract

The invention discloses a ribonucleic acid (RNA) extraction kit without deoxyribonucleic acid (DNA) residues and an RNA extraction method. The RNA extraction kit without the DNA residues comprises RNA eluent, DNA eluent, at least one DNA adsorption column which is matched with the RNA eluent and at least one RNA adsorption column which is matched with the DNA eluent. The RNA extraction method at least comprises the following two steps of: (A), eluting RNA on the DNA adsorption column by using the RNA eluent; and (B), eluting DNA on the RNA adsorption column by using the DNA eluent. The RNA extraction method is simple and quick; and the extracted RNA is high in purity, DNA residues are a few and cannot be seen by naked eyes under a common ultraviolet lamp.

Description

technical field [0001] The invention relates to RNA extraction, which belongs to the field of nucleic acid purification, in particular to an RNA extraction kit and an RNA extraction method without DNA residues. Background technique [0002] The extraction and purification of RNA in molecular biology research is a basic work, and it is necessary to extract and purify RNA in various clinical tests or basic scientific research. One of the difficulties in RNA extraction and purification is that DNA is often left behind. The low purity of RNA caused by residual DNA may have adverse effects on downstream experiments. [0003] At present, there are two main methods for removing DNA residues in the process of extracting and purifying RNA both at home and abroad: one is the guanidine isothiocyanate one-step method to extract RNA (Trizol method) and removes it by the method of phenol-chloroform organic phase separation; It is removed by DNase digestion. The disadvantages of the for...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 张茂
Owner BEIJING AIDLAB BIOTECH
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