CN108548710A - A kind of prefabricated examination powder and its application method for quickly preparing nucleic acid electrophoresis buffer solution based on boric acid - Google Patents

Abstract

The invention belongs to the field of biological analysis, and provides a prefabricated test powder for rapidly preparing nucleic acid electrophoresis buffer based on boric acid and a use method thereof. Its composition and weight are as follows: Based on the preparation of 1L one-time (1×) concentration of electrophoresis buffer, each prefabricated test powder contains 9.6-12g of Tris base, 4.89-6.11g of boric acid, and 0.186-0.744g of Na ₂ EDTA • 2H ₂ O, 3.40-3.67 g of KH ₂ PO ₄ . When in use, at room temperature, each portion of the test powder is thoroughly mixed with 1L of deionized water to obtain 1L of electrophoresis buffer with a concentration of 1×. When preparing the electrophoresis buffer, the present invention omits the steps of weighing multiple medicines and adjusting the pH value of the solution in the conventional method, so that the preparation process of the buffer is completed within 5-10 minutes, and the efficiency of the nucleic acid electrophoresis experiment is improved.

Description

A prefabricated test powder for rapidly preparing nucleic acid electrophoresis buffer based on boric acid and its application method

technical field

The invention belongs to the field of biological analysis, and provides a prefabricated test powder for rapid preparation of nucleic acid electrophoresis buffer in biochemical or molecular biology experimental analysis and a method for using the same.

Background technique

Electrophoresis detection plays an important role in the biochemical and molecular biological analysis of nucleic acids. It is one of the routine methods for nucleic acid quality and concentration detection, and an important basis for analyzing the size, mass concentration and purity of DNA and RNA fragments. Traditional nucleic acid electrophoresis mainly uses acetate buffer system TAE (Tris-Acetic acid-EDTA) or borate buffer system TBE (Tris-Boric acid-EDTA). Compared with TAE buffer, TBE buffer has better nucleic acid stability and longer electrophoretic life, but whether it is TAE or TBE buffer, it needs to be prepared by the experimenter, including weighing a number of drugs and adjusting the pH value, etc. steps, wasting a lot of manpower and time. It is urgent to develop a simple, fast and accurate method for preparing electrophoresis buffer with fixed concentration and pH value to improve the overall efficiency of electrophoresis experiments.

Aiming at the above experimental requirements and insufficient methods, the present invention develops a prefabricated test powder for rapidly preparing nucleic acid electrophoresis buffer, which is an improved TBE buffer, using Tris base, boric acid, Na ₂ EDTA, and KH ₂ PO ₄ The main component is mixed with deionized water in a certain proportion, and the nucleic acid electrophoresis buffer can be quickly prepared within 5-10 minutes, reducing the preparation time for electrophoresis experiments and improving the efficiency of electrophoresis experiments.

Contents of the invention

Aiming at the problems existing in the prior art, the present invention provides a prefabricated test powder for rapidly preparing nucleic acid electrophoresis buffer based on boric acid. The main components of the test powder include Tris base (trishydroxymethylaminomethane), boric acid (H ₃ BO ₃ ), Na ₂ EDTA (disodium ethylenediaminetetraacetic acid), and KH ₂ PO ₄ (potassium dihydrogen phosphate) . The electrophoresis buffer prepared by using the test powder is an improved system of the traditional TBE (Tris-Boric acid-EDTA), that is, borate buffer.

In order to achieve the above object, technical scheme of the present invention is as follows:

A prefabricated test powder for rapid preparation of nucleic acid electrophoresis buffer based on boric acid. Its composition and weight are as follows: Based on the preparation of 1L of one-time (1×) concentration of electrophoresis buffer, each prefabricated test powder contains 9.6-12g of Tris base , 4.89-6.11 g of boric acid, 0.186-0.744 g of Na ₂ EDTA·2H ₂ O, 3.40-3.67 g of KH ₂ PO ₄ . The concentration of each component in the thus prepared electrophoresis buffer is: Tris base 80-100mmol/L, boric acid 80-100mmol/L, Na ₂ EDTA 0.5-2mmol/L, KH ₂ PO ₄ 25-27mmol/L, pH The value is stable between 7.8-8.2. All materials in the test powder are mixed thoroughly and stored or sealed in plastic containers or bags in loose powder or tablet form. The Na ₂ EDTA·2H ₂ O and KH ₂ PO ₄ can be replaced by other Na ₂ EDTA containing different crystal water, but the mass concentration of Na ₂ EDTA and KH ₂ PO ₄ remains unchanged.

The described electrophoresis buffer based on the preparation of 1 L double (1×) concentration, the components and weight of the prefabricated test powder are preferably: each prefabricated test powder contains 10.8g of Tris base, 5.5g of boric acid, 0.744g Na ₂ EDTA·2H ₂ O, 3.53g of KH ₂ PO ₄ ; the concentration of each component in the electrophoresis buffer thus prepared is: Tris base 90mmol/L, boric acid 90mmol/L, Na ₂ EDTA 1mmol/L, KH ₂ PO ₄ 26mmol/L, buffer pH value between 8.0-8.1.

A kind of use method of the prefabricated test powder of fast preparation nucleic acid electrophoresis buffer based on boric acid is as follows:

At room temperature, each part of the test powder was fully mixed with 1L of deionized water to obtain 1L of electrophoresis buffer with a concentration of 1×. This process does not need to weigh the test powder, and does not need to adjust the pH value of the buffer solution, and the operation is simple and fast.

A kind of preparation method of the prefabricated test powder of fast preparation nucleic acid electrophoresis buffer based on boric acid is as follows:

The prefabricated test powder can be prepared on an enlarged or reduced scale. Based on the mass fraction of each component in the 1L 1× concentration electrophoresis buffer test powder, and according to the product of the target buffer volume and concentration, each component in the reference is calculated. The magnification or reduction of the mass of the components. After weighing and mixing the components uniformly, they are stored or packaged in plastic containers or bags in the form of loose powder or compressed tablets. For example: if equipped with 1L 10× concentration of electrophoresis buffer test powder, the mass of each component in the standard shall be amplified to 10 times; if equipped with 5L of 0.5× concentration of electrophoresis buffer test powder, the mass of each component in the standard shall be amplified to 2.5 times.

The prefabricated test powder can also be prepared as a whole, weigh and uniformly mix the components according to the mass ratio of the components, and store them in plastic containers or bags. When preparing the electrophoresis buffer, according to the product of the concentration and volume of the buffer to be prepared, first calculate the mass of the prefabricated test powder required, then weigh the prefabricated test powder, and finally mix it fully with the deionized water of the volume of the buffer to be prepared to prepare Electrophoresis buffer was obtained.

Compared with prior art, the beneficial effect of the present invention is:

The prefabricated test powder provided by the present invention is an improved TBE buffer solution, and the use process of the prefabricated test powder provided is simple and fast, using Tris alkali, boric acid, Na ₂ EDTA, and KH ₂ PO ₄ as main components, and deionized After the water is mixed and miscible, the steps of weighing multiple drugs and adjusting the pH value of the solution in the conventional method are omitted, so that the buffer preparation process can be completed within 5-10 minutes, and the nucleic acid electrophoresis buffer can be quickly prepared, reducing the preparation time for electrophoresis experiments , improve the efficiency of electrophoresis experiments.

Detailed ways

The present invention will be further described below in conjunction with specific examples.

Example 1

Prepare prefabricated test powder based on the preparation of 1L 1× concentration nucleic acid electrophoresis buffer, each test powder contains 10.8g of Tris base, 5.5g of boric acid, 0.744g of Na ₂ EDTA·2H ₂ O, 3.53g of KH ₂ PO ₄ . Take a portion of the test powder and mix it thoroughly with 1L deionized water to obtain a 1× concentration electrophoresis buffer (Tris base 90mmol/L, boric acid 90mmol/L, Na ₂ EDTA 2mmol/L, KH ₂ PO ₄ 26mmol/L, pH value 8.02), after 5 hours of electrophoresis, the pH value of the positive electrode in the electrophoresis tank was 8.0, the pH value of the negative electrode was 8.4, the nucleic acid sample was stable, and the ultraviolet imaging was good.

Example 2

Slightly adjust the formula of the test powder in Example 1. Based on the preparation of 1L 1× concentration of electrophoresis buffer, each pre-made test powder contains 10.8g of Tris base, 5.5g of boric acid, and 0.372g of Na ₂ EDTA·2H ₂ O. 3.4 g _{of KH2PO4} . Take a portion of the test powder and mix it thoroughly with 1L deionized water to obtain a 1× concentration electrophoresis buffer (Tris base 90mmol/L, boric acid 90mmol/L, Na ₂ EDTA 1mmol/L, KH ₂ PO ₄ 25mmol/L, pH value 8.03), after 5 hours of electrophoresis, the pH value of the positive electrode in the electrophoresis tank was 8.1, the pH value of the negative electrode was 8.3, the nucleic acid sample was stable, and the ultraviolet imaging was good.

The above-described embodiments only express the implementation manner of the present invention, but can not be interpreted as the limitation to the scope of patent of the present invention, should point out, for those skilled in the art, under the premise of not departing from the concept of the present invention , and several modifications and improvements can also be made, which all belong to the protection scope of the present invention.

Claims (4)

1. a kind of prefabricated examination powder for quickly preparing nucleic acid electrophoresis buffer solution based on boric acid, which is characterized in that the prefabricated examination Each constituent and weight are as follows in powder：Based on the electrophoretic buffer for preparing mono- times of (1 ×) concentration of 1L, every part of prefabricated examination powder contains The Na of the Tris alkali of 9.6-12g, the boric acid of 4.89-6.11g, 0.186-0.744g₂EDTA·2H₂O, 3.40-3.67g KH₂PO₄；Thus each constituent is a concentration of in the electrophoretic buffer prepared：Tris alkali 80-100mmol/L, boric acid 80- 100mmol/L、Na₂EDTA 0.5-2mmol/L, KH₂PO₄25-27mmol/L, pH stable is between 7.8-8.2；It tries in powder All materials are thoroughly mixed, and are stored with loose powder or compression forms.

2. a kind of prefabricated examination powder for quickly preparing nucleic acid electrophoresis buffer solution based on boric acid according to claim 1, special Sign is that described based on the electrophoretic buffer for preparing mono- times of (1 ×) concentration of 1L, each constituent and weight of prefabricated examination powder are excellent It is selected as：The Na of Tris alkali containing 10.8g, the boric acid of 5.5g, 0.744g in every part of prefabricated examination powder₂EDTA·2H₂O, 3.53g KH₂PO₄；Thus each constituent is a concentration of in the electrophoretic buffer prepared：Tris alkali 90mmol/L, boric acid 90mmol/L, Na₂EDTA 1mmol/L, KH₂PO₄26mmol/L, pH of cushioning fluid is between 8.0-8.1.

3. a kind of prefabricated examination powder for quickly preparing nucleic acid electrophoresis buffer solution based on boric acid according to claim 1 or 2, It is characterized in that, the Na₂EDTA·2H₂O and KH₂PO₄It can be by other Na containing the different crystallizations water₂EDTA is replaced, it is desirable that Na₂EDTA and KH₂PO₄Mass concentration remain unchanged.

4. claims 1 or 2 or a kind of 3 any prefabricated examination powder for quickly preparing nucleic acid electrophoresis buffer solution based on boric acid Application method, it is characterised in that following steps：At room temperature, the examination powder of every portion and 1L deionized waters are sufficiently mixed mutually It is molten to get to the electrophoretic buffer of 1 × concentration of 1L.