KR20020029477A - Method for gene extraction by one step - Google Patents

Abstract

PURPOSE: A process for extracting a gene by using one step is provided, thereby simply and rapidly extracting the DNA gene directly used for PCR. CONSTITUTION: The process for extracting a gene from a tissue or a cell of an animal by using one step comprises the steps of: chopping the tissue or cell of animal and inserting the chopped tissue or cell into 100 microliter of the lysis buffer reagent; reacting the lysis buffer reagent at 90 deg.C for 20 minutes; centrifuging the resulting solution at 12,000 rpm and room temperature for 5 minutes to extract DNA gene and recovering the supernatant; and amplifying 1 microliter of the supernatant by using PCR method.

Description

Gene extraction method by one step {METHOD FOR GENE EXTRACTION BY ONE STEP}

The present invention relates to a method for extracting a gene by one step. More specifically, the present invention relates to a technique that enables PCR to be immediately performed by extracting DNA genes from animal tissues or cells in a single step.

In biotechnology, genetic engineering is the basis of the basics. Among them, the method of extracting DNA is the most basic process. Until now, only DNA extracted through various steps has been amplified without any major problem in PCR.

Next, a series of steps for extracting DNA genes through several steps and amplifying the genes by PCR method will be described.

<Step 1>

; Remove part of the animal's tissue or cells.

<Step 2>

; Using a lysis buffer, a portion of tissue or cells removed from the animal is crushed and reacted so that genes such as DNA, RNA, and protein are dissolved in the buffer solution.

<3rd course>

; Various genes dissolved in the buffer solution are washed thoroughly with phenol-chloroform solution.

<Step 4>

; Only DNA genes are selectively extracted using 100% alcohol or 100% iso-propanol solution.

<Step 5>

; Finally, a series of experiments are terminated by amplifying the desired DAN gene by PCR.

As described above, the conventional method was required to extract a DNA gene through several steps from the tissue or cells of the animal and was able to detect the desired gene by the PCR method, which consumed considerable time and expense. In addition, there was a disadvantage in that only one sample should be tested at a time.

In other words, if PCR is performed directly on the extracted DNA gene using the Lysis buffer used in the conventional method, PCR is smoothed by inhibiting the action of the reagent used in the PCR of the reagent in the Lysis buffer. It was not possible to amplify the desired genes by PCR only after several steps.

Accordingly, the present invention is to solve the above problems, an object of the present invention is to significantly extract the existing gene extraction method to facilitate the extraction of DNA genes from animal tissue or cells in one step. It is to provide a method of extracting genes in one step.

As a technical idea for achieving the above object of the present invention

A part of animal tissue or cells is removed from the lysine buffer reagent and crushed. The lysine buffer reagent is reacted for a predetermined time, the gene is extracted through centrifugation, and then PCR is performed immediately using a predetermined amount of supernatant. A method of extracting a gene by one step is provided.

Hereinafter, a method of extracting a gene by one step according to the present invention will be described in detail.

First, a part of an animal tissue or cell is removed and crushed into 100 μl of Lysis buffer reagent, and the DNA gene is extracted by centrifugation after reacting the Lysis buffer reagent at 90 ° C. for 20 minutes.

At this time, the centrifugation process is carried out for 5 minutes at room temperature at a speed of 12000rpm.

Subsequently, a series of experiments according to the present invention are completed by selectively amplifying a desired gene by PCR using 1 μl of supernatant.

As described above, according to the present invention, it is possible to easily extract DNA genes from animal tissues or cells in one step by improving the conventional gene extraction method.

In other words, the conventional gene extraction method was difficult to amplify a desired gene by PCR because it affects the PCR buffer in PCR, but by using the improved gene extraction method according to the present invention without any problem by PCR In one step, the desired gene can be amplified by PCR.

As described above, according to the gene extraction method according to one step of the present invention, the following effects are obtained.

First, when PCR is performed immediately on DNA extracted from animal tissues using the existing gene extraction method, PCR is not smoothly performed by inhibiting the action of reagents used in PCR when the reagents contained in the lysis buffer are PCR. After several steps, the desired gene could be amplified by PCR.

However, according to the gene extraction method of the present invention, it is possible not only to make the lysis buffer smoothly, but also to directly use the extracted DNA gene in PCR in one step in the experiment.

Secondly, the range that can be tested at the same time was very limited because of the drawbacks of time-consuming and experimental steps when extracting DNA genes and performing PCR by conventional methods.

However, according to the gene extraction method by one step according to the present invention, not only the time can be shortened, but the test method is simplified, and thus a way to open a large-scale experiment can be opened.

Claims (4)

A part of animal tissue or cells is removed from the lysine buffer reagent, and then crushed into small pieces. The lysine buffer reagent is reacted for a predetermined time, the gene is extracted through centrifugation, and the PCR is performed immediately using a predetermined amount of supernatant. Gene extraction method by one step comprising a. The method of claim 1, wherein when the Lysis buffer reagent is 100 μl, the Lysis buffer reagent is reacted at 90 ° C. for 20 minutes. The method of claim 1, wherein the centrifugation process before gene extraction is performed at room temperature at 12000 rpm for 5 minutes. The method of claim 1, wherein the desired gene is amplified by PCR using 1 μl of the supernatant.