产品说明

一般描述

The High Pure RNA Tissue Kit is designed for the purification of total, intact RNA from tissue samples, free of any contaminating DNA. Low to medium throughput RNA isolation. Nucleic acids bind to the surface of the glass fleece in the presence of a chaotropic salt (guanidine HCl). This allows the High Pure Filter Tube to specifically immobilize nucleic acids (both DNA and RNA) while they are freed of contaminants. For RNA isolation, the binding conditions can be optimized to ensure immobilization of all the RNA. High pure RNA isolation kit can rapidly isolate intact total RNA from a broad range of research sample materials, including cultured cells, mammalian blood, white blood cells (WBCs), yeast, and bacteria. It rapidly isolates total RNA from mammalian tissues such as mouse liver, spleen, lung, and heart.

Capacity: The High Pure Spin Filter Tubes hold up to 700 μL sample volume.
Sample Material: Solid tissue (e.g., mouse liver, spleen, lung, heart): 1 - 25 mg.

Note: Sample size depends on the method used to prepare the tissue; larger samples (>10 mg) should be processed via rotor-stator homogenization.

应用

The High Pure RNA Tissue Kit rapidly isolates total RNA from mammalian tissues such as mouse liver, spleen, lung, and heart. The isolated RNA can be used in many downstream applications:

• RT-PCR
• Northern blotting
• RACE
• Primer extension
• Differential display
• cDNA library construction
• RNase protection assay
• In vitro translation

特点和优势

• Prepare RNA samples in approximately 30 minutes.
• Obtain concentrated RNA that is suitable for downstream applications.
• Minimize RNA loss with a kit that removes contaminants without precipitation or other handling steps that degrade RNA.
• Eliminate the use of hazardous organic compounds such as cesium chloride, phenol, chloroform, and ethidium bromide.

组分

• 裂解/结合缓冲液
• 脱氧核糖核酸酶ⅰ，冻干
• 脱氧核糖核酸酶孵育缓冲液
• 洗涤缓冲液I
• 洗涤缓冲液II
• 洗脱缓冲液
• 高纯纺丝过滤管(含玻璃纤维绒)
• 收集管

质量

• 组织在裂解/结合缓冲液中破碎和均质化，并如所述进行纯化。
• 核糖核酸产量是通过测量260纳米的光密度来确定的。
• 完整性和大小分布通过变性琼脂糖凝胶中核糖体核糖核酸的条带模式来检测。
• 以逆转录酶MuLV和₁₅为引物，在第一链合成中使用10 & # 956；L的核糖核酸洗脱液。在随后的聚合酶链反应中，使用扩增高保真聚合酶链反应系统和甘油醛3-磷酸脱氢酶(GAPDH)的特异性引物，获得了预期的983 bp的扩增产物。
• 通过聚合酶链反应进行检测，而没有预先的逆转录反应，结果为不存在污染性的脱氧核糖核酸；没有获得扩增产物。

制备说明

Tissue samples are disrupted and homogenized in the presence of a chaotropic salt (guanidine HCL) that inactivates RNases. The homogenate is then applied to the glass fiber fleece in a High Pure Spin Filter Tube. Under the buffer conditions used in the procedure, all nucleic acids bind to the glass fiber fleece, while contaminating substances (salts, proteins, and other cellular contaminants) do not. DNA in the preparation is digested with DNase I directly on the filter. Brief wash-and-spin steps readily remove the digested DNA fragments and other cellular contaminants. The remaining purified RNA is then eluted in a small volume of low-salt buffer.

其他说明

High Pure Technology and Silica Adsorption Kits
For life science research only. Not for use in diagnostic procedures.
图1：不同均质化程序的比较。使用各种程序均质化小鼠肝脏。用高纯核糖核酸组织试剂盒从每种匀浆中纯化总核糖核酸。分离的核糖核酸的等分试样用特异性针对谷氨酸脱羧酶基因区域的引物进行反向转录。通过凝胶电泳分析cDNA产物。

泳道1：均质化：Ultra Turrax；产量：1.9 & # 956；g/mg组织；A 260/ A 280 ： 2.0
泳道2：均质化：电动一次性塑料杵；产量：3 & # 956；g/mg组织；A 260 /A 280 ： 2.0
泳道3：均质化：研钵和杵，20 G针头；产量：1.5 & # 956；g/mg组织；A 260 /A 280 ： 2.0
泳道4：均质化：手动一次性塑料杵，20 G针头；产量：1.8 & # 956；g/mg组织；A 260 /A 280 ： 2.0
泳道5：均质化：手动一次性塑料杵；产量：3.4 & # 956；g/mg组织；A 260 /A 280 ： 2.0
泳道6：均质化：小珠涡旋；产量：3 & # 956；g/mg组织；A 260 /A 280 ： 2.0
泳道7：分子量标记

产品性质

安全信息

象形图
警示用语：	Danger
危险声明	H302 + H332 - H315 - H317 - H319 - H334
预防措施声明	P261 - P264 - P280 - P284 - P304 + P340 + P312 - P342 + P311
危险分类	Acute Tox. 4 Inhalation - Acute Tox. 4 Oral - Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - Skin Sens. 1
储存分类代码	12 - Non Combustible Liquids
WGK	WGK 1
闪点(F)	does not flash
闪点(C)	does not flash