基本描述

英文名称

RNApure Virus Kit

储存温度

室温

运输条件

常规运输

产品介绍

本试剂盒采用可以特异性结合病毒RNA的吸附柱和独特的缓冲液系统，适用于从血清、血浆、尿液、脑脊液等无细胞体液及细胞培养上清液中分离病毒RNA。病毒 RNA特异性地结合到硅基质膜上，而污染物则流过该膜。通过两次高效洗涤完全去除蛋白质等杂质，然后用无RNase的水或试剂盒提供的RNase-Free Water洗脱高纯度的病毒RNA。由本试剂盒提取的病毒RNA可直接用于RT-PCR、Real-time RT-PCR和印迹分析等实验。

R666005	Component	50 T	Storage
R666005A	Buffer GL	15 mL	RT
R666005B	Buffer RW1	40 mL	RT
R666005C	Buffer RW2（concentrate）	11 mL	RT
R666005D	Proteinase K	12.5 mg	RT
R666005E	Proteinase K Storage Buffer	1.25 mL	RT
R666005F	RNase-Free Water	10 mL	RT
R666005G	Spin Columns RS with Collection Tubes	50 sets	RT
R666005H	RNase-Free Centrifuge Tubes（1.5 mL）	50 EA	RT

自备试剂：无水乙醇，0.9% NaCl。

实验前准备及重要注意事项

1．向Proteinase K中加入1.25 ml Proteinase K Storage Buffer使其溶解，-20℃保存。配制好的Proteinase K勿长时间室温放置，避免反复冻融，以免影响其活性。

2．预防RNase污染，应注意以下几方面：

1）使用无RNase的塑料制品和枪头，避免交叉污染。

2）玻璃器皿应在使用前于180℃高温下干烤4小时，塑料器皿可在0.5 M NaOH中浸泡10分钟，用水彻底冲洗后高压灭菌。

3）配制溶液应使用无RNase的水。

4）操作人员戴一次性口罩和手套，实验过程中要勤换手套。

3．血清或血浆避免反复冻融导致蛋白变性或产生沉淀，减少病毒滴度进而影响提取病毒核酸的产量。

4．第一次使用前应按照试剂瓶标签的说明先在Buffer RW2中加入无水乙醇。

5．Buffer GL如果产生沉淀，可在56℃加热使其溶解后室温放置。

6．所有离心步骤若无特殊说明均在室温下进行，且所有操作步骤动作要迅速。

操作步骤

1．室温下取200 μl血清或血浆加到1.5 ml离心管（自备）中。注意：不足200 μl可以加入0.9 % NaCl （客户自备）补足。

2．向上步溶液中加入20 μl Proteinase K，混匀。

3．加入200 μl Buffer GL，涡旋震荡15秒。注意：不要直接把Proteinase K加到Buffer GL中。

4．56℃ 孵育15分钟，短暂离心，将管壁上的溶液收集到管底。

5．加入250 μl无水乙醇，涡旋震荡15秒，室温孵育5分钟，短暂离心，将管壁上的溶液收集到管底。

6．将步骤5得溶液全部加入到已装入收集管的吸附柱（Spin Columns RS）中，若一次不能将全部溶液加入吸附柱中，请分两次转入，12,000 rpm（~13,400×g）离心1 分钟，倒掉收集管中的废液，将吸附柱重新放回收集管中。

7．向吸附柱中加入500 μl Buffer RW1，12,000 rpm离心1分钟，倒掉收集管中的废液，将吸附柱重新放回收集管中。

8．向吸附柱中加入500 μl Buffer RW2（使用前检查是否加入无水乙醇），12,000 rpm 离心1分钟，倒掉收集管中的废液，将吸附柱重新放回收集管中。

9．向吸附柱中加入500 μl无水乙醇，12,000 rpm离心1分钟，倒掉收集管中的废液，将吸附柱重新放回收集管中。

10.12,000 rpm 离心3分钟，倒掉收集管中的废液。将吸附柱置于室温数分钟，以彻底晾干。

注意：

1）这一步的目的是将吸附柱中残余乙醇去除，乙醇的残留会影响后续的酶促反应（酶切、 PCR 等）。

2）推荐步骤：将吸附柱放入一个新的1.5 ml离心管（自备）中，打开管盖，56℃烘箱孵育3分钟，使吸附柱的膜彻底干燥。

11.将吸附柱置于一个新的无RNase离心管中，向吸附柱的中间部位悬空加入20-50 μl RNase-Free Water，室温放置5分钟，12,000 rpm离心1分钟，收集RNA溶液， -70℃保存RNA，防止降解。

注意：

1） RNase-Free Water体积不应小于20 μl，体积过小影响回收率。

2）如果要提高RNA的产量，可用20-50 μl新的RNase-Free Water重复步骤11。

3）如果要提高RNA浓度，可将得到的溶液重新加入到吸附柱中，重复步骤11。

This reagent kit uses an adsorption column that can specifically bind to viral RNA and a unique buffer system, suitable for isolating viral RNA from cell-free body fluids such as serum, plasma, urine, cerebrospinal fluid, and cell culture supernatants. The viral RNA specifically binds to the silicon substrate membrane, and pollutants flow through the membrane. Completely remove impurities such as proteins through two efficient washes, and then wash high-purity viral RNA with RNase free water or RNase Free Water provided by the reagent kit. The virus RNA extracted by this kit can be directly used for experiments such as RT-PCR, Real time RT-PCR, and Western blot analysis.

R666005	Component	50 T	Storage
R666005A	Buffer GL	15 mL	RT
R666005B	Buffer RW1	40 mL	RT
R666005C	Buffer RW2（concentrate）	11 mL	RT
R666005D	Proteinase K	12.5 mg	RT
R666005E	Proteinase K Storage Buffer	1.25 mL	RT
R666005F	RNase-Free Water	10 mL	RT
R666005G	Spin Columns RS with Collection Tubes	50 sets	RT
R666005H	RNase-Free Centrifuge Tubes（1.5 mL）	50 EA	RT

Self prepared reagent: anhydrous ethanol, 0.9% NaCl.
Preparation and important precautions before the experiment
1. Add 1.25 ml of Protein K Storage Buffer to Protein K to dissolve it and store at -20 ℃. The prepared Protein K should not be left at room temperature for a long time to avoid repeated freeze-thaw cycles, which may affect its activity.
2. To prevent RNase pollution, attention should be paid to the following aspects:
1) Use RNase free plastic products and gun heads to avoid cross contamination.
2) Glassware should be dry baked at a high temperature of 180 ℃ for 4 hours before use, while plastic containers can be soaked in 0.5 M NaOH for 10 minutes, thoroughly rinsed with water, and then sterilized under high pressure.
3) Prepare the solution using water without RNase.
4) Operators should wear disposable masks and gloves, and change gloves frequently during the experiment.
3. Serum or plasma should avoid repeated freeze-thaw cycles that may cause protein denaturation or precipitation, reduce viral titers, and thus affect the yield of extracted viral nucleic acids.
4. Before the first use, anhydrous ethanol should be added to Buffer RW2 according to the instructions on the reagent bottle label.
5. If buffer GL precipitates, it can be heated at 56 ℃ to dissolve and then placed at room temperature.
6. All centrifugation steps should be carried out at room temperature unless otherwise specified, and all operation steps should be carried out quickly.

Operation steps
1. Take 200 at room temperature μ Add serum or plasma to a 1.5 ml centrifuge tube (self provided). Attention: Less than 200 μ 0.9% NaCl (provided by the customer) can be added to make up for it.
2. Add 20 to the solution in the previous step μ Protein K, mix well.
3. Add 200 μ L Buffer GL, vortex oscillation for 15 seconds. Note: Do not directly add Protein K to Buffer GL.
4. Incubate at 56 ℃ for 15 minutes, briefly centrifuge, and collect the solution on the tube wall to the bottom of the tube.
5. Add 250 μ Anhydrous ethanol, vortex for 15 seconds, incubate at room temperature for 5 minutes, briefly centrifuge, and collect the solution from the tube wall to the bottom of the tube.
6. Add all the solution obtained in step 5 to the Spin Columns RS that have been loaded into the collection tube. If it is not possible to add all the solution to the adsorption column at once, please transfer it in two batches, centrifuge at 12000 rpm (~13400 × g) for 1 minute, discard the waste liquid in the collection tube, and put the adsorption column back into the collection tube.
7. Add 500 to the adsorption column μ Centrifuge at 12000 rpm for 1 minute, discard the waste liquid from the collection tube, and place the adsorption column back into the collection tube.
8. Add 500 to the adsorption column μ Buffer RW2 (check if anhydrous ethanol is added before use), centrifuge at 12000 rpm for 1 minute, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.
9. Add 500 to the adsorption column μ Centrifuge anhydrous ethanol at 12000 rpm for 1 minute, discard the waste liquid from the collection tube, and place the adsorption column back into the collection tube.
10. Centrifuge at 12000 rpm for 3 minutes and discard the waste liquid from the collection tube. Place the adsorption column at room temperature for a few minutes to thoroughly air dry.

Attention:
1) The purpose of this step is to remove residual ethanol from the adsorption column, which will affect subsequent enzymatic reactions (such as enzyme digestion, PCR, etc.).
2) Recommended steps: Place the adsorption column into a new 1.5 ml centrifuge tube (provided), open the tube cover, and incubate in a 56 ℃ oven for 3 minutes to thoroughly dry the membrane of the adsorption column.
11. Place the adsorption column in a new RNase free centrifuge tube and add 20-50 to the middle of the adsorption column in the air μ Place RNase Free Water at room temperature for 5 minutes, centrifuge at 12000 rpm for 1 minute, collect RNA solution, and store RNA at -70 ℃ to prevent degradation.
Attention:
1) The volume of RNase Free Water should not be less than 20 μ l. Small volume affects the recovery rate.
2) If you want to increase RNA production, you can use 20-50 μ Repeat step 11 for the new RNase Free Water.
3) If you want to increase the RNA concentration, you can add the obtained solution back to the adsorption column and repeat step 11.

安全和危险性（GHS）

SDS英文版

技术规格说明书

技术规格说明书

r666005_rnapure virus kit.pdf

质检证书（CoA,COO,BSE/TSE 和分析图谱)

C of A & Other Certificates(BSE/TSE, COO):

输入批号以搜索分析图谱:

溶液计算器

摩尔浓度计算器

计算溶液所需的质量、体积或浓度。

质量

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浓度

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分子量

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稀释计算器

计算配制储备溶液所需的稀释度。

浓度1（C1）

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体积 1 (V1)

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浓度 2 (C2)

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复溶计算器

复溶计算器允许您快速计算试剂的体积，以复溶小瓶。只需输入试剂的质量和目标浓度，计算器将确定其余部分。

体积（添加到小瓶中）

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质量（小瓶）

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所需的复溶浓度

病毒RNA提取试剂盒

库存信息

基本描述

安全和危险性（GHS）

技术规格说明书

质检证书（CoA,COO,BSE/TSE 和分析图谱)

溶液计算器

摩尔浓度计算器

稀释计算器

复溶计算器