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CN110343696A - A kind of quick, safe and nontoxic RNA extraction method - Google Patents

A kind of quick, safe and nontoxic RNA extraction method Download PDF

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CN110343696A
CN110343696A CN201910555079.7A CN201910555079A CN110343696A CN 110343696 A CN110343696 A CN 110343696A CN 201910555079 A CN201910555079 A CN 201910555079A CN 110343696 A CN110343696 A CN 110343696A
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顾月清
张行
陈红红
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China Pharmaceutical University
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Abstract

本发明公开了一种快速、安全、无毒的RNA提取方法,先在样本中加入裂解液和蛋白酶K工作液,使样本裂解释放出DNA/RNA,再采用氯仿抽提得到核酸水溶液,然后加入核酸助沉剂,向得到的固体核酸中,加入DNase1消化液去除DNA残留,之后加入DNase1终止液使DNase1失活,即可。本发明的方法可以高效去除样本中的蛋白质、脂类、无机盐等杂质,提取的核酸纯度高,片段完整,提取过程快速安全无毒。

The invention discloses a fast, safe and non-toxic RNA extraction method. Firstly, a lysate and a proteinase K working solution are added to a sample, so that the sample is cleaved to release DNA/RNA, and then extracted with chloroform to obtain an aqueous nucleic acid solution, and then added As a nucleic acid precipitation aid, add DNase1 digestion solution to the obtained solid nucleic acid to remove DNA residues, and then add DNase1 stop solution to inactivate DNase1. The method of the invention can efficiently remove impurities such as proteins, lipids and inorganic salts in samples, and the extracted nucleic acid has high purity and complete fragments, and the extraction process is fast, safe and non-toxic.

Description

一种快速、安全、无毒的RNA提取方法A fast, safe and non-toxic RNA extraction method

技术领域technical field

本发明属于生物技术领域,具体涉及一种快速、安全、无毒的RNA提取方法。The invention belongs to the field of biotechnology, and in particular relates to a fast, safe and nontoxic RNA extraction method.

背景技术Background technique

分子生物学研究中提取纯化RNA是一项基础性工作,各种临床检测或者基础科研中都需要提取纯化RNA。RNA提取纯化的难点在于:一是样本裂解不完全,RNA没有完全释放出来,二是由于DNA残留严重导致RNA纯度不高,对下游使用RT-PCR一步法进行检测分析造成不利影响。The extraction and purification of RNA in molecular biology research is a basic work, and it is necessary to extract and purify RNA in various clinical tests or basic scientific research. The difficulty of RNA extraction and purification lies in: firstly, the sample is not completely lysed, and the RNA is not completely released; secondly, the RNA purity is not high due to the serious DNA residue, which has an adverse effect on the downstream one-step detection and analysis by RT-PCR.

目前国内外提取纯化RNA的方法主要有异硫氰酸胍一步法提取RNA(Trizol法)、硫氰酸胍/酚法、酚/SDS法、盐酸胍法、CTAB法、柱提取法等,这些方法所用到的苯酚和胍盐的毒性都很大,在通过苯酚氯仿有机分相的方法来去除DNA残留过程中会有大量的RNA损失,导致检测灵敏度低,约在1000 copies/mL左右,检测范围窄,一般在1.00E+03 copies/mL-1.00E+07 copies/mL 之间,对于高临床值(大于5.00E+07 copies/mL)和低临床值(小于1.00E+03 copies/mL)的样本无法检测到,而柱提取法操作步骤繁琐需频繁更换离心管,用时长,特异性差,成本高,不易操作。At present, the methods for extracting and purifying RNA at home and abroad mainly include guanidine isothiocyanate one-step method for RNA extraction (Trizol method), guanidine thiocyanate/phenol method, phenol/SDS method, guanidine hydrochloride method, CTAB method, column extraction method, etc. The phenol and guanidine salts used in the method are very toxic, and a large amount of RNA will be lost in the process of removing DNA residues by phenol-chloroform organic phase separation, resulting in low detection sensitivity, which is about 1000 copies/mL. Narrow range, generally between 1.00E+03 copies/mL-1.00E+07 copies/mL, for high clinical value (greater than 5.00E+07 copies/mL) and low clinical value (less than 1.00E+03 copies/mL ) samples cannot be detected, and the column extraction method is cumbersome and requires frequent replacement of centrifuge tubes, which takes a long time, has poor specificity, high cost, and is not easy to operate.

因此,目前迫切需要一种毒性小、样本裂解完全、DNA残留去除彻底、无RT-PCR抑制物的RNA提取方法,可以将提取的RNA直接用RT-PCR的一步法检测。Therefore, there is an urgent need for an RNA extraction method with low toxicity, complete sample lysis, complete removal of DNA residues, and no RT-PCR inhibitors, which can directly detect the extracted RNA by one-step RT-PCR.

发明内容Contents of the invention

本发明的目的是解决上述现有技术的不足,提供一种不使用苯酚、胍盐等毒性物质,的无DNA残留的RNA提取方法,并且能够有效解决样本裂解不完全,RNA释放不彻底以及RNA提取纯化中的DNA残留问题。The purpose of the present invention is to solve the above-mentioned deficiencies in the prior art, to provide a method for extracting RNA without DNA residues without using toxic substances such as phenol and guanidinium salts, and to effectively solve incomplete lysis of samples, incomplete release of RNA and RNA extraction. The problem of DNA residue in extraction and purification.

为了实现上述目的,本发明采用以下技术手段:In order to achieve the above object, the present invention adopts the following technical means:

一种快速、安全、无毒的RNA提取方法,包括以下步骤:A fast, safe and non-toxic RNA extraction method comprising the following steps:

步骤1,在样本中加入裂解液和蛋白酶K工作液,使样本裂解释放出DNA/RNA;Step 1, add lysate and proteinase K working solution to the sample, so that the sample is lysed to release DNA/RNA;

步骤2,向步骤1得到的裂解混合液中加入氯仿抽提液,混匀、静置、离心,取上清,得到核酸水溶液;Step 2, adding chloroform extract to the lysis mixture obtained in step 1, mixing, standing, centrifuging, and taking the supernatant to obtain an aqueous nucleic acid solution;

步骤3,向步骤2得到的核酸水溶液中加入核酸助沉剂,混匀、静置、离心,弃去上清,得到固体核酸;Step 3, adding a nucleic acid sedimentation aid to the nucleic acid aqueous solution obtained in step 2, mixing, standing, centrifuging, discarding the supernatant to obtain solid nucleic acid;

步骤4,向步骤3得到的固体核酸中,加入DNase1消化液去除DNA残留,然后加入DNase1终止液使DNase1失活,即可。Step 4: Add DNase1 digestion solution to the solid nucleic acid obtained in step 3 to remove DNA residues, and then add DNase1 stop solution to inactivate DNase1.

进一步地,步骤1中蛋白酶K工作液的终浓度为187-281 μg/mL。Further, the final concentration of proteinase K working solution in step 1 is 187-281 μg/mL.

蛋白酶K工作液用于蛋白质的裂解,一方面可以裂解细胞,另一方面可以裂解蛋白释放核酸。Proteinase K working solution is used for protein cleavage, on the one hand, it can lyse cells, and on the other hand, it can lyse proteins to release nucleic acids.

进一步地,所述裂解液包括:β-巯基乙醇1%-2% v/v、Proclin 300 0.2%-2% v/v、50-55 mM HEPES酸、50-55 mM HEPES-Na盐、10-15 mM EDTA、1.2-1.5 M氯化锂、187-220 mM十二烷基硫酸锂;裂解液pH值为4.5-6.8。Further, the lysate includes: β-mercaptoethanol 1%-2% v/v, Proclin 300 0.2%-2% v/v, 50-55 mM HEPES acid, 50-55 mM HEPES-Na salt, 10 -15 mM EDTA, 1.2-1.5 M lithium chloride, 187-220 mM lithium dodecyl sulfate; the pH of the lysate is 4.5-6.8.

在裂解液中:EDTA作为变性剂,通过破坏细胞膜参与样本细胞的裂解;proclin300作为一种防腐剂,用于保存样本;β-巯基乙醇作为内源性RNase A的抑制剂,保护RNA不被降解。In the lysate: EDTA is used as a denaturant to participate in the lysis of sample cells by destroying the cell membrane; proclin300 is used as a preservative to preserve samples; β-mercaptoethanol is used as an inhibitor of endogenous RNase A to protect RNA from degradation .

进一步地,所述核酸助沉剂为75%乙醇和异丙醇按体积比2:1混合的混合液。Further, the nucleic acid precipitation aid is a mixture of 75% ethanol and isopropanol in a volume ratio of 2:1.

进一步地,所述DNase1终止液为20mM 二水合EDTA二钠溶液。Further, the DNase1 stop solution is 20 mM disodium EDTA dihydrate solution.

进一步地,步骤1中加入蛋白酶K工作液后的裂解条件为58 ℃-65 ℃、20min。Further, the cleavage conditions after adding proteinase K working solution in step 1 are 58°C-65°C, 20min.

进一步地,步骤4中DNase1消化液的反应条件是37 ℃水浴孵育30 min。Further, the reaction condition of the DNase1 digestion solution in step 4 is to incubate in a water bath at 37°C for 30 min.

进一步地,步骤4中DNase1终止液的反应条件是65 ℃水浴孵育10 min。Further, the reaction condition of the DNase1 stop solution in step 4 is to incubate in a water bath at 65 °C for 10 min.

上述方法在组织样本、拭子洗液、分泌物、细菌、粪便、细胞样本RNA提取中的应用。Application of the above method in RNA extraction of tissue samples, swab washing fluid, secretions, bacteria, feces, and cell samples.

本发明提取的RNA纯度和浓度可以用OneDrop OD1000进行检测,0D260/OD280比值是衡量蛋白质污染程度的指标,高质量的RNA,0D260/OD280读数在1.8-2.1之间,本发明提取的RNA的0D260/OD280典型的比值为1.8-2.1。The purity and concentration of the RNA extracted by the present invention can be detected by OneDrop OD1000, and the OD260/OD280 ratio is an index to measure the degree of protein contamination. For high-quality RNA, the OD260/OD280 reading is between 1.8-2.1, and the OD260 of the RNA extracted by the present invention is A typical ratio of /OD280 is 1.8-2.1.

本发明提取的RNA完整性可用普通琼脂糖凝胶电泳(电泳条件:胶浓度1.2%;1xTBE电泳液;120 V,20 min)进行检测。由于细胞中70%-80%的RNA为rRNA,电泳后UV下可以看到明显的rRNA条带。The integrity of the RNA extracted by the present invention can be detected by ordinary agarose gel electrophoresis (electrophoresis conditions: gel concentration 1.2%; 1xTBE electrophoresis solution; 120 V, 20 min). Since 70%-80% of RNA in cells is rRNA, obvious rRNA bands can be seen under UV after electrophoresis.

本发明的方法可以高效去除样本中的蛋白质、脂类、无机盐等杂质,提取的核酸纯度高,片段完整,提取过程快速安全无毒。The method of the invention can efficiently remove impurities such as proteins, lipids and inorganic salts in samples, and the extracted nucleic acid has high purity and complete fragments, and the extraction process is fast, safe and non-toxic.

附图说明Description of drawings

图1是Trizol法提取的大肠杆菌的总RNA的电泳图,泳道0为标记(Marker),泳道1、2、3是用南京鼎思生物科技有限公司的Trizol RNA提取试剂提取的大肠杆菌的总RNA,其中1、2、3是三个复孔。Figure 1 is the electrophoresis image of the total RNA of Escherichia coli extracted by the Trizol method, lane 0 is the marker (Marker), and lanes 1, 2, and 3 are the total RNA of Escherichia coli extracted with the Trizol RNA extraction reagent of Nanjing Dingsi Biotechnology Co., Ltd. RNA, where 1, 2, and 3 are three replicate wells.

图2是实施例1和实施例2提取得到的RNA的电泳图,泳道0为标记(Marker),泳道4是用南京鼎思生物科技有限公司的Trizol RNA提取试剂提取的Hela细胞的总RNA,泳道5、6是用本发明提取的Hela细胞的总RNA,泳道7、8是用本发明提取的大肠杆菌的总RNA。Figure 2 is the electrophoresis graph of the RNA extracted in Example 1 and Example 2. Lane 0 is the marker (Marker), and lane 4 is the total RNA of Hela cells extracted with Trizol RNA extraction reagent from Nanjing Dingsi Biotechnology Co., Ltd. Swimming lanes 5 and 6 are the total RNA of Hela cells extracted by the present invention, and swimming lanes 7 and 8 are the total RNA of Escherichia coli extracted by the present invention.

具体实施方式Detailed ways

下面结合具体实施例对本发明的技术方案作进一步说明。The technical solutions of the present invention will be further described below in conjunction with specific embodiments.

实施例1Example 1

提取Hela细胞的总RNAExtraction of total RNA from Hela cells

步骤1,取一管Hela细胞,1000 rpm,离心5 min,弃去上清,加入300 uL裂解液和3uL(20mg/ mL)蛋白酶K,裂解液涡旋混匀,于65 ℃孵育20 min,得到裂解物。Step 1. Take a tube of Hela cells, centrifuge at 1000 rpm for 5 min, discard the supernatant, add 300 uL of lysate and 3 uL (20 mg/mL) of proteinase K, vortex the lysate, and incubate at 65 °C for 20 min. to obtain a lysate.

所述裂解液含有β-巯基乙醇1%-2%(体积分数)、Proclin 300 0.2%-2%(体积分数)、 50-55 mM HEPES酸、50-55 mM HEPES-Na盐、10-15 mM EDTA、1.2-1.5 M氯化锂、187-220 mM 十二烷基硫酸锂;裂解液的制备方法是:称取EDTA 0.075 g,1.27 g氯化锂,0.298g Hepes酸,0.330 g Hepes-Na盐,1.27 g十二烷基硫酸锂,量取24.45 mL DEPC水,超声溶解后,加入50 uL proclin 300和500 uL β-巯基乙醇并混匀。The lysate contains β-mercaptoethanol 1%-2% (volume fraction), Proclin 300 0.2%-2% (volume fraction), 50-55 mM HEPES acid, 50-55 mM HEPES-Na salt, 10-15 mM EDTA, 1.2-1.5 M lithium chloride, 187-220 mM lithium dodecyl sulfate; the preparation method of the lysate is: weigh 0.075 g of EDTA, 1.27 g of lithium chloride, 0.298 g of Hepes acid, 0.330 g of Hepes- Na salt, 1.27 g lithium dodecyl sulfate, measure 24.45 mL DEPC water, after ultrasonic dissolution, add 50 uL proclin 300 and 500 uL β-mercaptoethanol and mix well.

步骤2,向裂解物里加入1/5体积(64 uL)的氯仿抽提液,上下颠倒15次,室温静置5min后,12000 g/min,离心15 min,上清即为核酸水溶液。Step 2: Add 1/5 volume (64 uL) of chloroform extract to the lysate, turn it upside down 15 times, let stand at room temperature for 5 minutes, centrifuge at 12000 g/min for 15 minutes, and the supernatant is the nucleic acid aqueous solution.

步骤3,取100 uL上清于1.5 mL无酶管中,加入1 mL核酸助沉剂,上下颠倒15次,室温静置10 min后,7500 g/min,离心10 min,沉淀即为固体核酸。Step 3: Take 100 uL of supernatant in a 1.5 mL enzyme-free tube, add 1 mL of nucleic acid sedimentation aid, turn it upside down 15 times, let stand at room temperature for 10 minutes, then centrifuge at 7500 g/min for 10 minutes, and the precipitate is solid nucleic acid .

步骤4,弃去上清,倒置在吸水纸上10 min,加入70 uL DEPC水,20 ul DNase1反应液,10 uL DNase1消化液,涡旋混匀后于37 ℃孵育30 min,加入10 uL DNase1消化终止液涡旋混匀,于65 ℃孵育10 min,冷却至室温后加1 mL核酸助沉剂,上下颠倒15次,室温静置10 min后,7500 g/min,离心10 min,倒置在吸水纸上10 min,加10 uL DEPC水涡旋混匀得到RNA溶液。Step 4, discard the supernatant, invert on absorbent paper for 10 min, add 70 uL DEPC water, 20 uL DNase1 reaction solution, 10 uL DNase1 digestion solution, vortex and mix well, incubate at 37 °C for 30 min, add 10 uL DNase1 Vortex and mix the digestion termination solution, incubate at 65 °C for 10 min, add 1 mL of nucleic acid precipitation aid after cooling to room temperature, invert up and down 15 times, let stand at room temperature for 10 min, centrifuge at 7500 g/min for 10 min, and invert on After 10 min on absorbent paper, add 10 uL DEPC water and vortex to mix to obtain RNA solution.

琼脂糖凝胶电泳检测,结果如图2所示。从图2可以看出采用本发明方法提取的Hela细胞的RNA残留DNA几乎没有,并且RNA的完整性明显好于Trizol法提取的RNA。Agarose gel electrophoresis detection, the results are shown in Figure 2. It can be seen from Fig. 2 that the RNA of the Hela cells extracted by the method of the present invention has almost no residual DNA, and the integrity of the RNA is obviously better than that of the RNA extracted by the Trizol method.

实施例2Example 2

提取大肠杆菌的总RNAExtraction of total RNA from Escherichia coli

步骤1,取一管大肠杆菌菌液,8000 rpm,离心3 min,弃去上清,加入300 uL裂解液和3uL(20mg/ mL)蛋白酶K,裂解液涡旋混匀,于65 ℃孵育20 min,得到裂解物。Step 1. Take a tube of E. coli liquid, centrifuge at 8000 rpm for 3 min, discard the supernatant, add 300 uL of lysate and 3 uL (20 mg/mL) proteinase K, vortex the lysate, and incubate at 65 °C for 20 min, to get the lysate.

所述裂解液含有β-巯基乙醇1%-2%(体积分数)、Proclin 300 0.2%-2%(体积分数)、 50-55 mM HEPES酸、50-55 mM HEPES-Na盐、10-15 mM EDTA、1.2-1.5 M氯化锂、187-220 mM 十二烷基硫酸锂;裂解液的制备方法是:称取EDTA 0.113 g,1.55 g氯化锂,0.328g Hepes酸,0.363 g Hepes-Na盐,1.5 g十二烷基硫酸锂,量取24.45 mL DEPC水,超声溶解后,加入500 uL proclin 300和250 uL β-巯基乙醇并混匀。The lysate contains β-mercaptoethanol 1%-2% (volume fraction), Proclin 300 0.2%-2% (volume fraction), 50-55 mM HEPES acid, 50-55 mM HEPES-Na salt, 10-15 mM EDTA, 1.2-1.5 M lithium chloride, 187-220 mM lithium dodecyl sulfate; the preparation method of the lysate is: weigh 0.113 g of EDTA, 1.55 g of lithium chloride, 0.328 g of Hepes acid, 0.363 g of Hepes- Na salt, 1.5 g lithium dodecyl sulfate, measure 24.45 mL DEPC water, after ultrasonic dissolution, add 500 uL proclin 300 and 250 uL β-mercaptoethanol and mix well.

步骤2,向裂解物里加入1/5体积(64 uL)的氯仿抽提液,上下颠倒15次,室温静置5min后,12000 g/min,离心15 min,上清即为核酸水溶液。Step 2: Add 1/5 volume (64 uL) of chloroform extract to the lysate, turn it upside down 15 times, let stand at room temperature for 5 minutes, centrifuge at 12000 g/min for 15 minutes, and the supernatant is the nucleic acid aqueous solution.

步骤3,取100 uL上清于1.5 mL无酶管中,加入1 mL核酸助沉剂,上下颠倒15次,室温静置10 min后,7500 g/min,离心10 min,沉淀即为固体核酸。Step 3: Take 100 uL of supernatant in a 1.5 mL enzyme-free tube, add 1 mL of nucleic acid sedimentation aid, turn it upside down 15 times, let stand at room temperature for 10 minutes, then centrifuge at 7500 g/min for 10 minutes, and the precipitate is solid nucleic acid .

步骤4,弃去上清,倒置在吸水纸上10 min,加入70 uL DEPC水,20 uL DNase1反应液,10 uL DNase1消化液,涡旋混匀后于37 ℃孵育30 min,加入10 uL DNase1消化终止液涡旋混匀,于65 ℃孵育10 min,冷却至室温后加1 mL核酸助沉剂,上下颠倒15次,室温静置10 min后,7500 g/min,离心10 min,倒置在吸水纸上10 min,加10 uL DEPC水涡旋混匀得到RNA溶液。Step 4, discard the supernatant, invert on absorbent paper for 10 min, add 70 uL DEPC water, 20 uL DNase1 reaction solution, 10 uL DNase1 digestion solution, vortex and mix well, incubate at 37 °C for 30 min, add 10 uL DNase1 Vortex and mix the digestion termination solution, incubate at 65 °C for 10 min, add 1 mL of nucleic acid precipitation aid after cooling to room temperature, invert up and down 15 times, let stand at room temperature for 10 min, centrifuge at 7500 g/min for 10 min, and invert on After 10 min on absorbent paper, add 10 uL DEPC water and vortex to mix to obtain RNA solution.

琼脂糖凝胶电泳检测,结果如图2所示。从图1和图2可以看出采用本发明方法提取的大肠杆菌的RNA残留DNA几乎没有,并且RNA的完整性明显好于Trizol法提取的RNA。Agarose gel electrophoresis detection, the results are shown in Figure 2. It can be seen from Fig. 1 and Fig. 2 that the Escherichia coli RNA extracted by the method of the present invention has almost no residual DNA, and the integrity of the RNA is obviously better than that of the RNA extracted by the Trizol method.

Claims (9)

1.一种快速、安全、无毒的RNA提取方法,其特征在于:包括以下步骤:1. A fast, safe, nontoxic RNA extraction method, characterized in that: comprises the following steps: 步骤1,在样本中加入裂解液和蛋白酶K工作液,使样本裂解释放出DNA/RNA;Step 1, add lysate and proteinase K working solution to the sample, so that the sample is lysed to release DNA/RNA; 步骤2,向步骤1得到的裂解混合液中加入氯仿抽提,混匀、静置、离心,取上清,得到核酸水溶液;Step 2, adding chloroform to the lysis mixture obtained in step 1 for extraction, mixing, standing, and centrifuging, and taking the supernatant to obtain an aqueous nucleic acid solution; 步骤3,向步骤2得到的核酸水溶液中加入核酸助沉剂,混匀、静置、离心,弃去上清,得到固体核酸;Step 3, adding a nucleic acid sedimentation aid to the nucleic acid aqueous solution obtained in step 2, mixing, standing, centrifuging, discarding the supernatant to obtain solid nucleic acid; 步骤4,向步骤3得到的固体核酸中,加入DNase1消化液去除DNA残留,然后加入DNase1终止液使DNase1失活,即可。Step 4: Add DNase1 digestion solution to the solid nucleic acid obtained in step 3 to remove DNA residues, and then add DNase1 stop solution to inactivate DNase1. 2.根据权利要求1所述的RNA提取方法,其特征在于:步骤1中蛋白酶K工作液的终浓度为187-281 μg/mL。2. The RNA extraction method according to claim 1, characterized in that: the final concentration of proteinase K working solution in step 1 is 187-281 μg/mL. 3.根据权利要求1所述的RNA提取方法,其特征在于:所述裂解液包括:β-巯基乙醇1%-2% v/v、Proclin 300 0.2%-2% v/v、 50-55 mM HEPES酸、50-55 mM HEPES-Na盐、10-15 mMEDTA、1.2-1.5 M氯化锂、187-220 mM 十二烷基硫酸锂;裂解液pH值为4.5-6.8。3. RNA extraction method according to claim 1, is characterized in that: described lysate comprises: β-mercaptoethanol 1%-2% v/v, Proclin 300 0.2%-2% v/v, 50-55 mM HEPES acid, 50-55 mM HEPES-Na salt, 10-15 mM EDTA, 1.2-1.5 M lithium chloride, 187-220 mM lithium dodecyl sulfate; the pH of the lysate is 4.5-6.8. 4.根据权利要求1所述的RNA提取方法,其特征在于:所述核酸助沉剂为75%乙醇和异丙醇按体积比2:1混合的混合液。4. The RNA extraction method according to claim 1, characterized in that: the nucleic acid sedimentation aid is a mixture of 75% ethanol and isopropanol at a volume ratio of 2:1. 5.根据权利要求1所述的RNA提取方法,其特征在于:所述DNase1终止液为20mM 二水合EDTA二钠溶液。5. The RNA extraction method according to claim 1, characterized in that: the DNase1 stop solution is 20mM dihydrate EDTA disodium solution. 6.根据权利要求1所述的RNA提取方法,其特征在于:步骤1中加入蛋白酶K工作液后的裂解条件为58 ℃-65 ℃、20min。6. The RNA extraction method according to claim 1, characterized in that: the lysis condition after adding proteinase K working solution in step 1 is 58°C-65°C, 20min. 7.根据权利要求1所述的RNA提取方法,其特征在于:步骤4中DNase1消化液的反应条件是37 ℃水浴孵育30 min。7. The RNA extraction method according to claim 1, characterized in that: the reaction condition of the DNase1 digestion solution in step 4 is to incubate in a water bath at 37°C for 30 min. 8.根据权利要求1所述的RNA提取方法,其特征在于:步骤4中DNase1终止液的反应条件是65 ℃水浴孵育10 min。8. The RNA extraction method according to claim 1, characterized in that: the reaction condition of the DNase1 stop solution in step 4 is to incubate in a water bath at 65°C for 10 min. 9.权利要求1所述的方法在组织样本、拭子洗液、分泌物、细菌、粪便、细胞样本RNA提取中的应用。9. the method for claim 1 is in the application of tissue sample, swab washing liquid, secretion, bacterium, feces, cell sample RNA extraction.
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Publication number Priority date Publication date Assignee Title
CN110904096A (en) * 2019-12-23 2020-03-24 广州湾区生物科技有限公司 Method for extracting RNA (ribonucleic acid) without DNA (deoxyribonucleic acid) by using inhibitor
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