基本描述

英文别名

TRIzol Reagent

稳定性与储存

Store at 2-8℃ for 12 months. Store in the dark.

英文名称

Trizol (Total RNA Extractor Reagent)

储存温度

2-8°C储存,避免反复冻融

运输条件

冰袋运输

产品介绍

产品介绍：

TRzol试剂是直接从细胞或组织中提取总RNA的试剂。它在破碎和裂解细胞时能保持RNA的完整性。加入氯仿替代物/氯仿后离心，样品分成水样层、中间层和有机层。RNA存在于水样层中。收集上面的的水样层后，RNA可以通过异丙醇沉淀来回收。无论是人、动物、植物还是细菌，该方法对少量及大量的组织和细胞均有较好的分离效果。TRzol试剂操作上的简单性允许同时处理多个样品。所有的操作可以在一小时内完成。TRzol抽提的总RNA能够避免DNA和蛋白的污染，可用于RNA印迹分析、斑点杂交、poly(A)+筛选、体外翻译、RNA酶保护分析和分子克隆。如果是用于 PCR，当两条引物位于单一外显子内时，建议用扩增级的DNase I来处理抽提的总RNA。TRzol试剂能促进不同种属不同分子量大小的多种RNA的析出。例如，从大鼠肝脏抽提的RNA经琼脂糖凝胶电泳并用溴化乙啶染色，可见许多介于7kb和15kb之间不连续的高分子量条带(mRNA和hnRNA成分)，两条优势核糖体的分子量约在5 kb (28S)和2 kb(18S)，低分子量RNA介于0.1和0.3kb之间(tRNA, 5S)。当抽提的RNA用TE稀释时其A260/A280比值≥1.8。

产品组分：

T669996	Component	100 ml	Storage
T669996A	TRzol	100 ml	2-8℃. Store in the dark.
T669996B	氯仿替代物	10 ml	2-8℃. Store in the dark.

注意事项

1. 从少量的组织(1~10mg)或细胞(102-104个)中分离RNA样品：往组织或细胞中加入800µl TRzol。待样品裂解后，加入氯仿替代物并进行步骤中的抽提操作。在用异丙醇沉淀RNA之前，加入5~10μg无RNA酶的glycogen作为水样层的载体。为降低其黏度在加入氯仿前用 26 号注射器抽吸两次以切断基因组DNA。Glycogen 会留在水样层中并和RNA共析出。在浓缩到 4mg/ml 之前它不会抑制逆转录反应第一链的合成也不会抑制 PCR。

2. 在匀浆化后和加入氯仿替代物（R749970）之前，样品可以在-60℃或者-70℃保存至少一个月。将 RNA 沉淀溶于 75%的乙醇在 2-8℃至少可以保存一周，在-5—-20℃下至少可保存一年。

3. 用 TRzol 抽提 RNA 时要戴手套和护眼罩。避免接触皮肤和衣服。在化学通风橱完成操作。避免呼吸道吸入。如无例外，室温保持在 15~30℃的条件下。

Product Introduction:

TRzol Reagent is a reagent designed for the direct extraction of total RNA from cells or tissues. It maintains the integrity of RNA during the disruption and lysis of cells. Upon the addition of a chloroform substitute/chloroform and subsequent centrifugation, the sample separates into an aqueous phase, an interphase, and an organic phase. RNA is present in the aqueous phase. After collecting the upper aqueous phase, RNA can be recovered by isopropanol precipitation. This method is effective for both small and large amounts of tissues and cells, regardless of whether they are from humans, animals, plants, or bacteria. The simplicity of TRzol Reagent allows for the simultaneous processing of multiple samples. All operations can be completed within one hour. Total RNA extracted with TRzol is free from DNA and protein contamination and can be used for RNA blotting analysis, dot hybridization, poly(A)+ selection, in vitro translation, RNase protection analysis, and molecular cloning. When used for PCR, it is recommended to treat the extracted total RNA with amplification-grade DNase I if both primers are located within a single exon. TRzol Reagent facilitates the precipitation of various types of RNA with different molecular weights across species. For example, RNA extracted from rat liver, when subjected to agarose gel electrophoresis and stained with ethidium bromide, shows numerous discontinuous high molecular weight bands between 7kb and 15kb (mRNA and hnRNA components), two major ribosomal bands with molecular weights of approximately 5kb (28S) and 2kb (18S), and low molecular weight RNA between 0.1 and 0.3kb (tRNA, 5S). When the extracted RNA is diluted with TE, its A260/A280 ratio is ≥1.8.

Product component:

T669996	Component	100 ml	Storage
T669996A	TRzol	100 ml	2-8℃. Store in the dark.
T669996B	Chloroform substitute	10 ml	2-8℃. Store in the dark.

Precautions:

1. Isolation of RNA samples from small amounts of tissue (1-10mg) or cells (10²-10⁴ cells): Add 800µl of TRzol to the tissue or cells. After the sample is lysed, add the chloroform substitute and proceed with the extraction steps. Before precipitating RNA with isopropanol, add 5-10µg of RNase-free glycogen as a carrier for the aqueous phase. To reduce viscosity, pass the sample through a 26-gauge needle twice before adding chloroform to shear genomic DNA. Glycogen will remain in the aqueous phase and co-precipitate with RNA. It will not inhibit the synthesis of the first strand in reverse transcription reactions or PCR until it is concentrated to 4mg/ml.

2. Sample storage: The sample can be stored at -60℃ or -70℃ for at least one month before homogenization and addition of the chloroform substitute (R749970). RNA precipitate dissolved in 75% ethanol can be stored at 2-8℃ for at least one week and at -5 to -20℃ for at least one year.

3. Safety: When extracting RNA with TRzol, wear gloves and eye protection. Avoid contact with skin and clothing. Perform the operation in a chemical fume hood. Avoid inhalation through the respiratory tract. Unless otherwise specified, maintain the room temperature between 15-30℃.