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.
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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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