基本描述

英文名称

Ultrapure RNA Kit

储存温度

2-8°C储存,避光,室温

运输条件

冰袋运输

产品介绍

产品内容

U670000	Component	50 T	200 T	Storage
U670000A	TRIzon Reagent	60 mL	2×110 mL	2-8℃. Protect from light
U670000B	TRIzon PaI ™	10 mL	2x20 mL	2-8℃. Protect from light
U670000C	Buffer RW1	40 mL	160 mL	RT
U670000D	Buffer RW2 (concentrate)	11 mL	50 mL	RT
U670000E	RNase-Free Water	10 mL	50 mL	RT
U670000F	Spin Columns RM with Collection Tubes	50 EA	200 EA	RT
U670000G	RNase-Free Centrifuge Tubes (1.5 mL)	50 EA	200 EA	RT

产品简介

本试剂盒是基于TRIzon改进后的柱式总RNA提取试剂盒，裂解液充分裂解并匀质化样本，采用独特的硅基质膜吸附技术，通过离心吸附柱在高盐状态下高效专一的结合溶液中的RNA，同时最大限度的有效除去蛋白质、无机盐离子及有机杂质等；可从动物组织、植物材料、各种微生物及培养细胞等样品中快速提取总RNA，每次可处理30-50 mg组织或5×10⁶细胞，可同时处理多个不同样品。本试剂盒提取得到的RNA可直接应用于RT-PCR、Northern Blot、Dot Blot、体外翻译等实验。

产品特点

• 纯度高：最大限度除去蛋白质等杂质，提取的RNA可直接用于下游各种实验。

• 提取量大：独特的裂解液配方，充分裂解细胞或组织，RNA提取量多至100 μg。

• 快速：步骤少，操作简单，节省时间。

• 兼容性强：适用于多种动植物组织和细胞RNA的提取。

自备试剂：

70%乙醇(无RNase水配制)、无水乙醇。

实验前准备及重要注意事项

1. 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。

2. 样品应避免反复冻融，否则影响RNA提取得率和质量。

3. 使用前若发现TRIzon Reagent有沉淀，可置于56℃水浴几分钟，即可溶解。

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

5. 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。

6. 若下游实验对DNA非常敏感，建议用不含RNase的DNase I 对 RNA进行处理。

使用方法

1. 样品处理

1a.植物组织：取新鲜植物组织在液氮中充分研磨或将植物组织剪碎后直接在TRIzon Reagent中迅速研磨，每30-50 mg组织加入1 mL TRIzon Reagent，混匀。

注意：样品体积不超过TRIzon Reagent体积的10%。

1b.动物组织：取新鲜或-70℃冻存的动物组织尽量剪碎，每30-50mg组织加入1 mL TRIzon Reagent，匀浆仪进行匀浆处理。或在液氮中研磨后加入TRIzon Reagent 1 mL混匀。

注意：样品体积一般不要超过TRIzon Reagent体积的10%。

1c.单层培养细胞：吸去培养液，可直接在培养板中加入适量TRIzon Reagent（每10 cm²面积需要1 mL TRIzon Reagent），用取样器反复吹打使细胞裂解。也可用胰蛋白酶处理后，将细胞溶液转移至RNase-Free的离心管中，300×g离心5 min，收集细胞沉淀，仔细吸除所有上清，加入TRIzon Reagent 1 mL混匀。

注意：

1） 收集细胞数量不要超过1×107。

2） TRIzon Reagent加量根据培养板面积决定，不是由细胞数决定。如果TRIzon Reagent加量不足，可能导致提取的RNA中有DNA污染。

3）收集细胞时一定要将细胞培养液去除干净，否则会导致裂解不完全，造成RNA的产量降低。

1d.细胞悬液：离心收集细胞。每5×10⁶—1×10⁷动物、植物和酵母细胞或每10⁷细菌细胞加入1 mL TRIzon Reagent。

注意：

1）加入TRIzon Reagent前不要洗涤细胞，以免RNA降解。

2）一些酵母和细菌细胞可能需要匀浆仪或液氮研磨处理。

1e.血液处理：直接取新鲜的血液，加入3倍体积的TRIzon Reagent（推荐0.25 mL全血加入0.75 mL TRIzon Reagent），充分振荡混匀。

1f.可选步骤：对于蛋白、脂肪、多糖或胞外物质含量高的样品，如肌肉组织、脂肪组织或植物的块茎，可以在匀浆处理后4℃，12,000 rpm（~13,400×g）离心10分钟以除去不溶物质，此时沉淀中含胞外物质、多糖和高分子量的DNA，而RNA存在于上清中。

2.样品中加入TRIzon Reagent后反复吹打几次，使样本充分裂解。室温放置5分钟，使蛋白核酸复合物完全分离。

3.每1 mL TRIzon Reagent加入200 μLTRIzon PaI ™，盖好管盖，剧烈振荡15秒，室温放置2分钟。

4.4℃ 12,000 rpm（~13,400×g）离心10分钟，此时样品分为三层：红色有机相，中间层和上层无色水相，RNA主要在上层水相中，将上层水相移到一个新的RNase- Free离心管（自备）中。

5.在得到的水相溶液中加入等体积的70%乙醇（无RNase水配制），颠倒混匀。

6.将上步所得溶液全部加入到已装入收集管的吸附柱（Spin Columns RM）中。若一次不能加完溶液，可分多次转入。12,000 rpm离心20秒，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700μL Buffer RW1，12,000 rpm离心20秒，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（使用前检查是否已加入无水乙醇），12,000 rpm离心20秒，倒掉收集管中的废液，将吸附柱重新放回收集管中。

9.重复步骤8。

10.12,000 rpm离心2 分钟，倒掉收集管中废液。将吸附柱置于室温数分钟，彻底晾干。

注意：这一步目的是将吸附柱中残余乙醇去除，乙醇残留会影响后续酶促反应（酶切、PCR等）。

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加入30-50 μL RNase-Free Water，室温放置1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 Wate体积不应小于30 μL，体积过小影响回收率。

2） 如果要提高RNA的产量，可用30-50 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。

Product content

U670000	Component	50 T	200 T	Storage
U670000A	TRIzon Reagent	60 mL	2×110 mL	2-8℃. Protect from light
U670000B	TRIzon PaI ™	10 mL	2x20 mL	2-8℃. Protect from light
U670000C	Buffer RW1	40 mL	160 mL	RT
U670000D	Buffer RW2 (concentrate)	11 mL	50 mL	RT
U670000E	RNase-Free Water	10 mL	50 mL	RT
U670000F	Spin Columns RM with Collection Tubes	50 EA	200 EA	RT
U670000G	RNase-Free Centrifuge Tubes (1.5 mL)	50 EA	200 EA	RT

Product Introduction

This kit is an improved column total RNA extraction kit based on TRIzon. The lysate is fully lysed and homogenized, and a unique silicon matrix membrane adsorption technology is used to efficiently and specifically bind RNA in a high salt state through a centrifugal adsorption column. At the same time, proteins, inorganic salt ions, and organic impurities are effectively removed to the maximum extent possible; Total RNA can be quickly extracted from animal tissues, plant materials, various microorganisms, and cultured cells. It can process 30-50 mg of tissue or 5 × 10 ⁶ cells each time, and can simultaneously process multiple different samples. The RNA extracted from this kit can be directly applied to RT-PCR Northern Blot、Dot Blot、 Experiments such as in vitro translation.

Product Features
• High purity: maximally removes impurities such as proteins, and the extracted RNA can be directly used for various downstream experiments.
• Large extraction capacity: With a unique lysis buffer formula, cells or tissues can be fully lysed, and RNA extraction can reach up to 100 μ g.
Quick: With fewer steps, simple operation, and time-saving.
Strong compatibility: suitable for extracting RNA from various animal and plant tissues and cells.

Self provided reagents:
70% ethanol (prepared without RNase water), anhydrous ethanol.

Preparation and important precautions before the experiment.

To prevent RNase contamination, attention should be paid to the following aspects:
1) Use RNase free plastic products and gun tips to avoid cross contamination.
2) Glassware should be dried and baked at a high temperature of 180 ℃ for 4 hours before use, while plastic utensils can be soaked in 0.5 M NaOH for 10 hours
Rinse thoroughly with water in minutes and then sterilize under high pressure.
3) The solution should be prepared using water without RNase.
4) Operators should wear disposable masks and gloves, and change gloves frequently during the experiment.
2. The sample should avoid repeated freezing and thawing, otherwise it will affect the yield and quality of RNA extraction.
If TRIzon Reagent precipitates before use, it can be dissolved in a 56 ℃ water bath for a few minutes.
Before the first use, anhydrous ethanol should be added to Buffer RW2 according to the instructions on the reagent bottle label.
5. All centrifugation steps should be carried out at room temperature unless otherwise specified, and all operation steps should be performed quickly.
6.If downstream experiments are highly sensitive to DNA, it is recommended to use DNase I without RNase to treat RNA.

Usage method:
1. Sample processing
1a. Plant tissue: Take fresh plant tissue and grind it thoroughly in liquid nitrogen or cut it into pieces and quickly grind it directly in TRIzon Reagent. Add 1 mL of TRIzon Reagent to every 30-50 mg of tissue and mix well.
Note: The sample volume should not exceed 10% of the TRIzon Reagent volume.
1b. Animal tissue: Fresh or -70 ℃ frozen animal tissue should be cut into pieces as much as possible, and 1 mL of TRIzon Reagent should be added to every 30-50mg of tissue for homogenization treatment using a homogenizer. Or grind in liquid nitrogen and add 1 mL of TRIzon Reagent to mix well.
Note: The sample volume should generally not exceed 10% of the TRIzon Reagent volume.
1c. Single layer cultured cells: Remove the culture medium and directly add an appropriate amount of TRIzon reagent (1 mL TRIzon reagent is required per 10 cm ² area) to the culture plate. Use a sampler to repeatedly blow and beat the cells to lyse. Alternatively, after treatment with trypsin, transfer the cell solution to a RNase Free centrifuge tube, centrifuge at 300 × g for 5 minutes, collect the cell pellet, carefully aspirate all supernatant, add 1 mL of TRIzon Reagent and mix well.
be careful:
1) The number of collected cells should not exceed 1 × 107.
2) The dosage of TRIzon Reagent is determined by the area of the culture plate, not by the number of cells. If the dosage of TRIzon Reagent is insufficient, it may lead to DNA contamination in the extracted RNA.
3) When collecting cells, it is necessary to remove the cell culture medium completely, otherwise incomplete lysis may occur, resulting in a decrease in RNA production.
3) When collecting cells, it is necessary to remove the cell culture medium completely, otherwise incomplete lysis may occur, resulting in a decrease in RNA production.
1d. Cell suspension: Collect cells by centrifugation. Add 1 mL TRIzon Reagent every 5 × 10 ⁶ -1 × 10 ⁷ of animal, plant, and yeast cells, or every 10 ⁷ of bacterial cells.
be careful:
1) Do not wash cells before adding TRIzon Reagent to prevent RNA degradation.
2) Some yeast and bacterial cells may require homogenization or liquid nitrogen grinding treatment.
1e. Blood processing: Take fresh blood directly and add 3 times the volume of TRIzon reagent (recommended to add 0.75 mL of TRIzon reagent to 0.25 mL of whole blood), shake thoroughly and mix well.
1f. Optional steps: For samples with high protein, fat, polysaccharide, or extracellular substance content, such as muscle tissue, adipose tissue, or plant tubers, centrifugation at 12000 rpm (~13400 × g) for 10 minutes after homogenization treatment can be performed to remove insoluble substances. At this time, the precipitate contains extracellular substances, polysaccharides, and high molecular weight DNA, while RNA is present in the supernatant.

2. After adding TRIzon Reagent to the sample, blow it repeatedly several times to fully lyse the sample. Leave at room temperature for 5 minutes to completely separate the protein nucleic acid complex.
3. Add 200 μ L TRIzon PaI to every 1 mL TRIzon Reagent ™， Cover the tube tightly, shake vigorously for 15 seconds, and let it stand at room temperature for 2 minutes.
Centrifuge at 4.4 ℃ and 12000 rpm (~13400 × g) for 10 minutes. At this time, the sample is divided into three layers: a red organic phase, a middle layer, and an upper colorless aqueous phase. RNA is mainly in the upper aqueous phase. Transfer the upper aqueous phase to a new RNase Free centrifuge tube (provided).
5. Add an equal volume of 70% ethanol (prepared without RNase water) to the obtained aqueous solution, invert and mix well.
6. Add all the solutions obtained in the previous step into the Spin Columns RM that have been loaded into the collection tube. If the solution cannot be added at once, it can be transferred in multiple times. Centrifuge at 12000 rpm for 20 seconds, discard the waste liquid from the collection tube, and place the adsorption column back into the recovery manifold.
7. Add 700 μ L Buffer RW1 to the adsorption column, centrifuge at 12000 rpm for 20 seconds, discard the waste liquid in the collection tube, and place the adsorption column back into the recovery header.
8. Add 500 μ L of Buffer RW2 to the adsorption column (check if anhydrous ethanol has been added before use), centrifuge at 12000 rpm for 20 seconds, discard the waste liquid in the collection tube, and place the adsorption column back into the recovery header.
9. Repeat step 8.
Centrifuge at 10.12000 rpm for 2 minutes and discard the waste liquid from the collection tube. Place the adsorption column at room temperature for a few minutes and let it dry thoroughly.
Attention: The purpose of this step is to remove residual ethanol from the adsorption column, which can affect subsequent enzymatic reactions (such as digestion, PCR, etc.).
11. Place the adsorption column in a new RNase free centrifuge tube, add 30-50 μ L RNase Free Water to the middle of the adsorption column, let it stand at room temperature for 1 minute, centrifuge at 12000 rpm for 1 minute, collect RNA solution, and store RNA at -70 ℃ to prevent degradation.
be careful:
1) The volume of RNase Free Wat should not be less than 30 μ L, as a small volume can affect the recovery rate.
2) If you want to increase RNA production, repeat step 11 with 30-50 μ L of 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

质量标准

质量标准

u670000_ultrapure rna 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

=

浓度

x

体积

x

分子量

g/mol

稀释计算器

计算配制储备溶液所需的稀释度。

浓度1（C1）

x

体积 1 (V1)

=

浓度 2 (C2)

x

体积 2 (V2)

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）

=

质量（小瓶）

÷

所需的复溶浓度

货号 (SKU)	包装规格	是否现货	价格	数量
U670000-200T	200T	期货
U670000-50T	50T	期货

超纯RNA提取试剂盒

库存信息

库存信息

基本描述

安全和危险性（GHS）

质量标准

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