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A method for rapidly extracting nucleic acid from biological samples

A biological sample and sample technology, applied in the field of rapid nucleic acid extraction, can solve the problem of not using Chelex-100 to extract RNA, etc., and achieve the effect of simple and easy method, short time, and elimination of adverse effects

Active Publication Date: 2014-10-29
苏州云泰生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] However, there is no report of using Chelex-100 to extract RNA at present. For this reason, the inventor proposed a simple, fast and effective method for extracting RNA from biological samples using Chelex-100

Method used

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  • A method for rapidly extracting nucleic acid from biological samples
  • A method for rapidly extracting nucleic acid from biological samples
  • A method for rapidly extracting nucleic acid from biological samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] 1. Two methods were used to extract DNA from paraffin-embedded tissue sections. Three samples were extracted by each method and compared in parallel.

[0043] 1) Extract DNA from paraffin-embedded tissue sections with a paraffin tissue section DNA extraction kit produced by Beijing Biotek Biotechnology Co., Ltd.

[0044] a. Take two pieces of 5um medical paraffin (model: model: BX12M311398, produced by Beijing Zhongxi Yuanda Technology Co., Ltd.) embedded tissue slices and put them into a 1.5mL centrifuge tube.

[0045] b. Add 1ml of xylene, shake and mix well, and bathe in water at 55°C for 10 minutes. Spin at 13000 for 5 minutes, and discard the supernatant.

[0046] c. Repeat step 2 one to three times.

[0047] d. Add 1ml of absolute ethanol, shake and mix. Spin at 13000 for 5 minutes, and discard the supernatant.

[0048] e. Repeat step 4 once.

[0049] f. Dry the settled specimens.

[0050] g. Add 200μl Buffer 1 and mix well.

[0051] h. Add 25μl Proteinase ...

Embodiment 2

[0085] 1. Place the fine spot sample in the aqueous solution of chelex-100, the concentration of the aqueous solution of chelex-100 is 20g / 100ml;

[0086] Heating at 2.95°C for 10 minutes;

[0087] 3. Put the extraction column into a centrifuge tube, add the liquid obtained in step 3 into the extraction column, centrifuge at 20,000 rpm for 20 seconds;

[0088] 4. Collect the separated liquid for PCR reaction.

[0089] Wherein, the extraction column is commercially available; the PCR reaction adopts the same conditions as in Example 1, and similar experimental results are obtained.

Embodiment 3

[0091] 1. Place the tissue fluid sample in the aqueous solution of chelex-100, the concentration of the aqueous solution of chelex-100 is 10g / 100ml;

[0092] 2. Heating at 100°C for 50 minutes;

[0093] 3. Put the extraction column into a centrifuge tube, add the liquid obtained in step 3 into the extraction column, centrifuge at 15,000 rpm for 30 seconds;

[0094] 4. Collect the separated liquid for PCR reaction.

[0095] Wherein, the extraction column is commercially available; the PCR reaction adopts the same conditions as in Example 1, and similar experimental results are obtained.

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Abstract

The present invention relates to a method for rapidly extracting nucleic acid from a biological sample. The method comprises: taking a biological sample, placing the sample in an aqueous solution of chelex-100 or a DEPC aqueous solution of chelex-100, and heating for 1-50 minutes at a temperature of 80-100 DEG C; placing an extraction column into a centrifuge tube, adding the resulting liquid to the extraction column, and centrifugating for 2-120 seconds at a rotation speed of 1000-20000 rotation / min; and taking the centrifugated liquid to carry out a PCR reaction or a RT-PCR reaction. According to the present invention, chelex-100 and a high temperature heating method are adopted to extract so as to eliminate a protease hydrolysis step, eliminate a paraffin section dewaxing step, remove an organic solvent xylene, and substantially shorten a time; and a retention membrane is placed in the extraction column so as to remove adverse effects on the PCR reaction or the RT-PCR reaction by the liquid paraffin and the chelex-100, and improve sensitivity of the PCR reaction or the RT-PCR reaction.

Description

technical field [0001] The invention relates to a method for rapidly extracting nucleic acid from biological samples. Background technique [0002] DNA molecule is the basic material of molecular biology research, and different extraction methods can be used to obtain DNA of varying quantity and quality according to different experimental purposes. Currently, there are several methods for extracting DNA: [0003] (1) Anionic detergent method: use detergents such as SDS or sodium xylene to denature proteins, and DNA can be directly extracted from biological materials. DNA and protein in cells are often combined by electrostatic attraction or coordination bonds. Since anionic detergents can destroy this valence bond, anionic detergents are commonly used to extract DNA, but anionic detergents often affect subsequent PCR reactions. [0004] (2) Water extraction method: Utilizing the nature of nucleic acid dissolving in water, after the tissue cells are broken, the RNA is remov...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
Inventor 熊慧谢立群陈嘉铮
Owner 苏州云泰生物医药科技有限公司
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