RNeasy Plus Mini Kit

For purification of up to 100 µg total RNA from cells/tissues using gDNA Eliminator columns

  • Unique gDNA Eliminator columns avoid the need for DNase
  • Efficient removal of genomic DNA
  • Highly reproducible yields of RNA in minutes
  • High-performance RNA for sensitive applications

The RNeasy Plus Mini Kit integrates fast, convenient purification of up to 100 µg RNA with effective elimination of genomic DNA. Cell or tissue lysates are spun through gDNA Eliminator spin columns to remove genomic DNA. Total RNA is then purified using RNeasy Mini spin columns. The kit can be automated on the QIAcube Connect. Tissue samples can be conveniently stabilized using RNAprotect Tissue Reagent or Allprotect Tissue Reagent, and efficiently disrupted using a TissueRuptor or TissueLyser system. For smaller samples, the RNeasy Plus Micro Kit (spin-column binding capacity of 45 µg RNA) is also available. Request a quote for a trial kit.

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产品 货号 目录价:
RNeasy Plus Mini Kit (50)
For 50 minipreps: RNeasy Mini Spin Columns, gDNA Eliminator Spin Columns, Collection Tubes, RNase-Free Water and Buffers
74134
3.870,00 kr
加入购物车
RNeasy Plus Mini Kit (250)
For 250 minipreps: RNeasy Mini Spin Columns, gDNA Eliminator Spin Columns, Collection Tubes, RNase-Free Water and Buffers
74136
17.035,00 kr
加入购物车

RNeasy Plus Mini Kit适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。


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High, reproducible RNA yields.|高效去除组织的基因组DNA。|RNeasy Plus实验流程。|高效去除细胞的基因组DNA。|可直接用于芯片分析的RNA。|
Total RNA was purified in duplicate from different amounts of Jurkat cells using the RNeasy Plus Mini Kit or a similar kit from Supplier AV. Real-time RT-PCR assays for β-actin were performed (40 cycles). The lower CT values with the RNeasy Kit demonstrate greater RNA yields. With the kit from Supplier AV, no transcript was detectable in RNA purified from 102 cells. |使用RNeasy Plus Mini Kit或其他供应商的试剂盒,从各种小鼠组织(每个样本10 mg)中纯化总RNA,重复两次。对c-jun进行real-time PCR分析,测定纯化RNA中DNA污染物的量。|利用较短的工作流程,只需不到30分钟即可完成RNA纯化并去除基因组DNA。|使用 [A] RNeasy Plus Mini Kit或者 [B] Supplier AV的整合基因组DNA去除步骤的RNA纯化试剂盒,从Jurkat细胞样本(每个样本1 x 106个细胞)中纯化总RNA。使用(+RT)或不使用(-RT)逆转录酶对β-actin进行两次重复的real-time RT-PCR分析。-RT曲线证明,使用RNeasy Plus Mini Kit纯化的RNA中几乎不含基因组DNA。|使用RNeasy Plus Mini Kit从1 x 106个HeLa细胞中纯化总RNA,重复两次。使用GeneChip IVT Labeling Kit从两个重复的RNA样本(各3.5 μg)中制备cRNA。在GeneChip Human Genome U133A probe芯片上分析cRNA样本(各15 μg)。散点图显示了两个样本间的相关性(Pearson相关系数 [r] 为0.996)。红色:两个样本中均存在的基因;蓝色:在一个样本中不存在或者处于临界状态的基因;黄色:在两个样本中均不存在或者处于临界状态的基因。|
性能
Purification of RNA from cells and tissues with the RNeasy Plus Mini procedure allows high, reproducible RNA yields and effective elimination of genomic DNA contamination for sensitive applications (see figures "Effective cell genomic DNA removal", "Effective tissue genomic DNA removal", "High, reproducible RNA yields", and "Array-ready RNA"). Total RNA with Agilent RIN values of close to 10 is routinely obtained from cultured cells.
原理

Cells and easy-to-lyse tissues are lysed and homogenized in highly denaturing guanidine-isothiocyanate–containing Buffer RLT Plus, which immediately inactivates RNases to ensure isolation of intact RNA. The lysate is then passed through a gDNA Eliminator spin column. This column, in combination with the high-salt buffer, selectively and efficiently removes genomic DNA. Ethanol is added to provide appropriate binding conditions for RNA, and the sample is applied to an RNeasy MinElute spin column. These specialized columns contain a silica membrane that specifically binds RNA from lysed cells.

操作流程

Total RNA is purified from up to 107 cells or 30 mg tissue. A short workflow enables RNA purification with genomic DNA removal in less than 25 minutes (see flowchart "RNeasy Plus procedure").  Samples are first lysed and homogenized. The lysate is passed through a gDNA Eliminator spin column, ethanol is added to the flow-through, and the sample is applied to an RNeasy MinElute spin column. RNA binds to the membrane and contaminants are washed away. High quality RNA is eluted in 30 µl, or more, of water.

Different protocols are available for different starting materials. The protocols differ mainly in the lysis and homogenization of the sample. Once the sample is applied to the gDNA Eliminator spin column, the protocols are similar. The procedure provides an enrichment for mRNA since most RNAs <200 nucleotides (such as rRNAs and tRNAs) are excluded. The RNeasy Plus procedure can be modified to allow the purification of total RNA containing small RNAs, such as miRNA, from cultured cells. 

When disrupting and homogenizing tissues in Buffer RLT Plus (supplied with the RNeasy Plus Mini Kit), excessive foaming may occur. This foaming is substantially reduced by adding Reagent DX (supplied separately) to Buffer RLT Plus before starting disruption and homogenization. Reagent DX has been carefully tested with the kit, and has no effect on RNA purity or on downstream applications.

应用

RNA purified using the RNeasy Plus Mini Kit is ideally suited for downstream applications that are sensitive to low amounts of DNA contamination, such as quantitative real-time RT-PCR. The purified RNA can also be used in other applications.

特点
参数
应用 PCR, qPCR, real-time RT-PCR, microarray
洗脱体积 30-50 µl
规格 Spin column
整合基因组DNA去除步骤 Yes
主要样本类型 Tissue, cells
处理 Manual (centrifugation)
纯化总RNA、miRNA、poly A+ mRNA、DNA或蛋白 Total RNA
样本量 30 mg (<700 µl)
技术 Silica technology
每次运行或制备时间 25 minutes
产量 Varies

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补充实验方案
2

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There are two protocols: follow Protocol 1 if you want to purify total RNA containing miRNA, or follow Protocol 2 if you want to purify small RNA (includes miRNA, 5S rRNA, and tRNA) and larger RNA (>200 nt) separately.
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学术海报
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参考文献
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图片
High, reproducible RNA yields.
High, reproducible RNA yields.
Total RNA was purified in duplicate from different amounts of Jurkat cells using the RNeasy Plus Mini Kit or a similar kit from Supplier AV. Real-time RT-PCR assays for β-actin were performed (40 cycles). The lower CT values with the RNeasy Kit demonstrate greater RNA yields. With the kit from Supplier AV, no transcript was detectable in RNA purified from 102 cells. 
Effective genomic DNA removal.
高效去除组织的基因组DNA。
使用RNeasy Plus Mini Kit或其他供应商的试剂盒,从各种小鼠组织(每个样本10 mg)中纯化总RNA,重复两次。对c-jun进行real-time PCR分析,测定纯化RNA中DNA污染物的量。
RNeasy Plus procedure.
RNeasy Plus实验流程。
利用较短的工作流程,只需不到30分钟即可完成RNA纯化并去除基因组DNA。
Effective elimination of genomic DNA contamination.
高效去除细胞的基因组DNA。
使用 [A] RNeasy Plus Mini Kit或者 [B] Supplier AV的整合基因组DNA去除步骤的RNA纯化试剂盒,从Jurkat细胞样本(每个样本1 x 106个细胞)中纯化总RNA。使用(+RT)或不使用(-RT)逆转录酶对β-actin进行两次重复的real-time RT-PCR分析。-RT曲线证明,使用RNeasy Plus Mini Kit纯化的RNA中几乎不含基因组DNA。
Array-ready RNA.
可直接用于芯片分析的RNA。
使用RNeasy Plus Mini Kit从1 x 106个HeLa细胞中纯化总RNA,重复两次。使用GeneChip IVT Labeling Kit从两个重复的RNA样本(各3.5 μg)中制备cRNA。在GeneChip Human Genome U133A probe芯片上分析cRNA样本(各15 μg)。散点图显示了两个样本间的相关性(Pearson相关系数 [r] 为0.996)。红色:两个样本中均存在的基因;蓝色:在一个样本中不存在或者处于临界状态的基因;黄色:在两个样本中均不存在或者处于临界状态的基因。