Rapid nucleic acid extraction kit and applications thereof

A kit and nucleic acid technology, applied in the field of biological nucleic acid detection, can solve the problems of cumbersome dewaxing process and harm to the body of the experimental operator, and achieve the effects of eliminating adverse effects, simple and easy methods, and shortening time.

Active Publication Date: 2012-12-05
上海源奇生物医药科技有限公司
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AI Technical Summary

Problems solved by technology

[0007] The traditional extraction of RNA from paraffin tissue needs to be dewaxed with xylene first, and then removed with absolute ethanol. The dewaxi...

Method used

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  • Rapid nucleic acid extraction kit and applications thereof
  • Rapid nucleic acid extraction kit and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1. Two methods were used to extract DNA from paraffin-embedded tissue sections. Three samples were extracted by each method and compared in parallel.

[0036] 1. Extract DNA from paraffin-embedded tissue sections with the Paraffin Tissue Section DNA Extraction Kit produced by Beijing Biotech Biotechnology Co., Ltd.:

[0037] a) Take two pieces of 5um medical paraffin (model: BX12M311398, produced by Beijing Zhongxi Yuanda Technology Co., Ltd.) embedded tissue slices and put them into a 1.5mL centrifuge tube.

[0038] 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.

[0039] c) Repeat step 2 one to three times.

[0040] d) Add 1ml of absolute ethanol, shake and mix. Spin at 13000 for 5 minutes, and discard the supernatant.

[0041] e) Repeat step 4 once.

[0042] f) Dry the settled specimens.

[0043] g) Add 200 μl Buffer 1 and mix well.

[0044] h) Add 25 μl Proteinase K...

Embodiment 2

[0078] 1. Use the kit of the present invention to quickly extract the RNA in the paraffin-embedded tissue section:

[0079] 1.1 Take 3 pieces of 5um medical paraffin (model: BX12M311398, produced by Beijing Zhongxi Yuanda Technology Co., Ltd.) to embed tissue sections, grind them on ice, and put them into a 1.5mL centrifuge tube;

[0080] 1.2 Place in 200ul solution 1, which is 10% chelex-100 DEPC aqueous solution;

[0081] 1. Heating at 390°C for 20 minutes;

[0082] 1.4 Put the extraction column tube into the centrifuge tube, add the liquid obtained in step 3 into the extraction column tube, and centrifuge at 12000 rpm for 30 seconds;

[0083] 1.5 Collect the separated liquid, take 5ul for RT-PCR reaction.

[0084] 2. Fluorescent quantitative RT-PCR detection of the concentration of the extracted RNA

[0085] Primers for human GAPDH and TaqMan fluorescent probes were selected, designed by Primer Express software, and synthesized by Shanghai Bioengineering Company (Sangong...

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Abstract

The invention belongs to the technical field of biological nucleic acid detection, and particularly relates to a rapid nucleic acid extraction kit and applications thereof. The kit of the invention comprises a grinding rod, an extraction liquid, an extraction column, a micro ultra-filter and a centrifuge tube, wherein the extraction liquid is a DEPC aqueous solution containing chelex-100. The invention further relates to a use method for the kit, and applications of the kit in extractions of DNA or RNA from pathological tissue paraffin section samples, samples of blood stains and semen stains, tissue fluid samples, cerebrospinal fluid samples, blood samples, serum samples, plasma samples, urine samples, hydrops samples, and tissue cell samples. According to the present invention, content and quality of the nucleic acid extracted by the kit of the present invention can well meet requirements of follow-up reactions of PCR, RT-PCR and sequencing. With the present invention, limitation of complexity and time consuming of nucleic acid extraction in the prior art are overcome well, and characteristics of safety and environmental protection are provided.

Description

technical field [0001] The invention belongs to the technical field of biological nucleic acid detection, in particular to a kit for rapidly extracting DNA or RNA from biological samples. Background technique [0002] Since the advent of PCR technology in the United States in 1985, it has been rapidly and widely used in molecular biology-related fields. Especially in recent years, with the concept of individualized medicine and its clinical promotion, RT-PCR is used to study patient samples. The difference in the expression levels of different genes in the test can be used to find out the change trend of key genes or prognostic genes, help doctors guide medication, and make RT-PCR enter clinical diagnosis and treatment. [0003] However, RT-PCR amplification must use RNA as a template to proceed. The current classic RNA extraction method is TRIZOL reagent, which directly extracts total RNA from cells or tissues. It preserves RNA integrity while disrupting and lysing cells....

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 熊慧谢立群陈嘉铮
Owner 上海源奇生物医药科技有限公司
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