CN102051357A - Super-micro deoxyribonucleic acid/ribonucleic acid electro-eluter - Google Patents

Abstract

The invention belongs to the field of life medicine science, and in particular relates to an ultra-miniature nucleic acid electric eluting instrument. It consists of an electrode, a miniature electroelution container, a sample gel block, an electroelution buffer and a cooling container, wherein: the electrode is inserted into the micro electroelution container, the sample gel block is located between the positive and negative electrodes, and the micro electroelution The de-vessel is located in the cooling vessel, and the miniature electroelution vessel is filled with electroelution buffer. Separating and purifying samples from gels is a common technical means and an important operation step in molecular biology experiments. At present, in the experiment of purifying nucleic acid and other molecules, commonly used instruments have disadvantages such as easy degradation of purified products, cumbersome operation, and high cost. In order to elute or recover nucleic acid (DNA and RNA) with high quality and low cost, especially RNA, the inventor has developed a miniature micro-nucleic acid electrowasher. In a micro-electrophoresis environment of less than 1ml, under low voltage (~50U) and in a short time (~20 minutes), trace nucleic acid molecules in PAGE gel can be effectively recovered, especially suitable for elution and recovery of easily degradable RNA samples.

Description

Ultraminiature Nucleic Acid Electroelution Apparatus

technical field

The invention belongs to the field of life medicine science, in particular to a super-mini DNA/RNA electro-eluter.

Background technique

Separating and purifying samples from gels is a common technical means and an important operation step in molecular biology experiments. The quality of sample separation and purification directly affects the downstream research work. At present, in the experiment of purifying nucleic acid and other molecules, commonly used instruments include Bio-rad’s Model 422 electro-Eluter and other instruments. This instrument is compact and applicable, and can meet the needs of most laboratories for separation and purification of biological macromolecules. However, when separating and purifying a small amount of nucleic acid samples, especially RNA samples that are easily degraded, the instrument is cumbersome to operate. First, even though the equipment is very sophisticated, it still has disadvantages such as large volume, many accessories, and many operating procedures for the small number of samples separated. It is not easy to completely inactivate RNase, and a small amount of contamination can reduce the sample RNA yield; Second, some parts of the equipment, such as Membrane Cap, etc., are improperly handled and installed or have air bubbles, which directly affect the efficiency of electroelution, or the electroelution process cannot be performed; third, the amount of electroelution buffer is large (700mL or so); Fourth, the cost of a complete set of equipment is relatively high. the

Contents of the invention

The purpose of the present invention is to provide an ultra-miniature nucleic acid electroelution instrument. the

The ultra-miniature nucleic acid electroelution instrument proposed by the present invention is composed of an electrode 1, a miniature electroelution container 2, a sample-containing gel block 3, an electroelution buffer 4, and a cooling container 5, wherein: the electrode 1 is inserted into the micro electroelution In the container 2, the gel block 3 containing the sample is located between the positive and negative electrodes 1, the miniature electroelution container 2 is located in the cooling container 5, and the miniature electroelution container 2 is filled with the electroelution buffer 4. the

In the present invention, the electric elution apparatus can be used in combination, that is, several miniature electric elution containers 2 are placed in the same cooling container 5 . the

The present invention can effectively recover trace nucleic acid molecules in PAGE gel under low voltage (~50U) and short time (~20 minutes) in a micro-electrophoresis environment less than 1ml, and is especially suitable for elution and recovery of easily degradable RNA samples . the

The present invention is characterized by: (1) extremely simple operation; (2) fast sample separation; (3) low operating cost; (4) suitable for the separation and purification of easily degradable RNA and other samples. the

Compared with similar products, the present invention has obvious advantages. Taking the model 422 electro-eluter of BIO-Rad Company as an example, the present invention has the following advantages:

Table 1. Comparison of equipment cost and consumables between model 422 electro-eluter and ultra-miniature electro-eluter

Description of drawings

Fig. 1 is a schematic diagram of the structure of the present invention. the

Fig. 2 is a structural diagram of Embodiment 2 of the present invention. the

Figure 3. Different amplification curves of miR-1a in samples eluted by model 422 and ultra-miniature electroelution instrument, among which: E1a: samples obtained after elution by ultra-miniature electroelution instrument (showing high sample yield) ; M1a: The sample volume is the same as the former, and the sample is obtained after elution with the model 422 elution instrument (showing a low sample yield). the

Figure 4. Different amplification curves of miRNA-1b in samples eluted by model 422 and ultra-miniature electroelution instrument, where: E1b: samples obtained after elution by ultra-miniature electroelution instrument (showing high sample yield) ; M1b: The same amount of sample as the former, the sample was obtained after elution using the model 422 elution instrument (showing a low sample yield). the

Figure 5. Different amplification curves of miRNA-2b in samples eluted by model 422 and ultra-miniature electroelution instrument, where: E2b: samples obtained after elution by ultra-miniature electroelution instrument (showing high sample yield) ; M2b: The same amount of sample as the former, the sample was obtained after elution using the model 422 elution instrument (showing a low sample yield). the

Numbers in the figure: 1 is an electrode; 2 is a miniature electric elution container; 3 is a gel block containing a sample; 4 is an electric elution buffer; 5 is a cooling container. the

Detailed ways

The present invention is further illustrated below by way of examples. the

Example 1:

As shown in Figure 1, the gel block 3 containing the sample is placed in the miniature electroelution container 2, and the electroelution buffer 4 has been housed in the container; the positive and negative electrodes 1 are inserted into the electroelution buffer 4, respectively placed in the container containing Both sides of the sample gel block 3; the entire miniature electroelution container is equipped with an external cooling container 5 to prevent the system from overheating; after power is applied, the RNA or DNA sample can swim out of the sample gel block 3 and enter the electroelution buffer 4 in. High-yield samples can be obtained by collecting electroelution buffer 4.

Embodiment 2: As shown in Figure 2, it shows that 4 samples are separated at the same time, and this device can increase the number of samples as required. The gel block 3 containing the sample is placed in the miniature electroelution container 2, and the electroelution buffer 4 is already installed in the container; the positive and negative electrodes 1 are inserted into the electroelution buffer 4, and placed on both sides of the gel block 3 containing the sample ; Place 4 miniature electric elution containers 2 in the same cooling container 5 to prevent the system from overheating; after power is applied, RNA or DNA samples can swim out of the sample-containing gel block 3 and enter the electroelution buffer 4 middle. Collect 4 electroelution buffers 4 separately to obtain high-yield samples. the

Carry out mass comparison to the elution sample with the model 422 electro-eluter electric elution apparatus of BIO-Rad company and the present invention:

Use the above two types of electroelution devices to elute the same RNA samples in the PAGE gel respectively. After electroelution, use Real-time PCR to detect the above three small RNAs in the electroelution samples, namely miR-1a, miR-1b and miR For the content of -2b, see Figure 3, Figure 4 and Figure 5, and compare the sample yields after elution from the two devices.

In Figure 3, E1a: sample eluted by ultra-miniature electroelution instrument, M1a: sample eluted by model 422, the sample E1a obtained after elution by ultra-miniature electroelution instrument was amplified by real-time PCR, and the number of cycles was less than that of model 422 The sample M1a obtained by the type electroeluter indicates that the concentration of the former sample is higher than that of the latter. the

Figure 3-5 shows that in the samples eluted by the ultra-miniature electroelution instrument, the small RNA content is higher than that of the Model 422 electroelution instrument, suggesting that the ultra-miniature electroelution instrument can recover samples efficiently, conveniently, and with high yield . the

Claims (2)

1. superminiature nov nucleic acid elution instrument, by electrode (1), miniature electroelution container (2), contain sample blob of viscose (3), electroelution damping fluid (4) and cooling vessel (5) and form, it is characterized in that electrode (1) inserts in the miniature electroelution container (2), containing sample blob of viscose (3) is positioned between the positive and negative electrode (1), miniature electroelution container (2) is positioned at cooling vessel (5), is full of electroelution damping fluid (4) in the miniature electroelution container (2).

2. superminiature nov nucleic acid elution instrument according to claim 1 is characterized in that described electroelution instrument is used in combination, and promptly several miniature electroelution containers (2) are positioned in the same cooling vessel (5).

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