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RNeasy Lipid Tissue Mini Kit

从富含脂肪的组织和其他类型组织中纯化至多100 µg的总RNA

  • 优化的裂解条件,用于富含脂肪的组织和其他组织
  • 高产量总RNA,无苯酚污染
  • 获得高品质RNA,适合多种下游应用
RNeasy Lipid Tissue Mini Kit包含QIAzol Lysis Reagent和RNeasy离心柱,前者用于裂解脂肪和其他类型组织样本,后者用于纯化至多100 µg高品质RNA。该试剂盒可在QIAcube全自动核酸纯化仪上全自动化完成。使用RNAlater RNA Stabilization Reagent(仅适用非脂肪组织)或Allprotect Tissue Reagent可方便的稳定组织样本,使用TissueRuptor或TissueLyser体系可有效地破碎样本。处理更大的样本可使用RNeasy Lipid Tissue Midi Kit(离心柱的结合能力为1 mg RNA)。

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Cat No./ID: 74804
RNeasy Lipid Tissue Mini Kit (50)
50 RNeasy Mini Spin Columns, Collection Tubes (1.5 ml and 2 ml), QIAzol Lysis Reagent, RNase-free Reagents and Buffers

RNeasy Lipid Tissue 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。


Product Details

9
RNeasy Lipid Tissue Mini procedure.

The convenient RNeasy Lipid Tissue protocol integrates QIAzol phenol/guanidine-based lysis with silica-membrane RNeasy RNA isolation for high yields of total RNA. The combination of organic extraction and chaotropic disruption contributes to the high lysis efficiency of QIAzol Lysis Reagent, enabling use of larger amounts of fatty tissues in the RNeasy procedure. RNA partitions to the upper, aqueous phase, while DNA partitions to the interphase and proteins to the lower, organic phase or the interphase. Ethanol is added to the aqueous phase and the sample is applied to the RNeasy spin column.

9
Real-time analysis of high-quality RNA.

Total RNA was isolated from 100 mg rat brain or adipose tissues using the RNeasy Lipid Tissue Mini Kit. Real-time, quantitative RT-PCR was carried out using QuantiTect Probe RT-PCR Kit with the indicated amounts of total RNA (in duplicate) and primers and probe specific for the c-jun or alpha-actin genes.

9
High yields of RNA without phenol carryover.
RNA was isolated from 10 mg rat brain tissue using the RNeasy Lipid Tissue Mini Kit (RNeasy Lipid), a standard silica-gel-membrane procedure (Std. silica), or a phenol/guanidine-based reagent (PhOH/Gdn), following supplier's instruction. [A] Formaldehyde agarose gel analysis shows high yields of RNA using the RNeasy Lipid Tissue Mini Kit. Using the RNeasy and other silica-based methods, small RNAs (such as 5.8S rRNA, 5S rRNA, and tRNAs) are selectively excluded. [B] Absorbance spectrum shows contaminants and phenol when using the phenol/guanidine-based reagent.
Performance
RNeasy Lipid Tissue操作流程将QIAzol裂解方法与RNeasy RNA分离方法结合,能从富含脂肪的组织中得到高产量总RNA,10 mg脑组织RNA的平均产量为10 µg、10 mg动物脂肪组织RNA的平均产量为2.4 µg。RNeasy Lipid Tissue Kits可有效的纯化高品质总RNA,无苯酚残留(参见"High-quality RNA" 和 "High yields of RNA without phenol carryover")。从从脂肪组织得到的RNA适用于包括real-time RT-PCR(参见"Real-time analysis" )等下游应用。
Principle

RNeasy Lipid Tissue Kits适用于富含脂肪的组织,如脑和脂肪组织。方便RNeasy Lipid Tissue实验方案结合了优化的苯酚/胍的裂解方法以及经验证的RNeasy硅胶膜纯化方法,能纯化得到高品质的总RNA。QIAzol Lysis Reagent结合了有机提取与离液破碎的方法,具有很高的裂解效率。RNeasy技术适用于大于200 nt RNA,因此RNA产物不包括5S rRNA、tRNA或其他小分子量RNAs,产物占细胞内总RNA的15–20%。

Procedure
10–100 mg组织样本在QIAzol Lysis Reagent中均质化。加入氯仿,匀浆离心后分为水相和有机相。吸取上层水相后加入乙醇以提供合适的结合条件。然后将样品加入RNeasy离心柱,至多100 µg的总RNA与硅胶膜结合,苯酚和其他污染物被高效洗去。使用30–100 µl不含RNase的纯水即可洗脱得到高品质RNA(参见"RNeasy Lipid Tissue Mini procedure)。
Applications

RNeasy Lipid Tissue Kits可简单有效的纯化高品质RNA,得到的RNA适用于多种下游应用,包括芯片分析和real-time RT-PCR。

Specifications

Features
Specifications
应用 PCR, real-time PCR, microarray
洗脱体积 30–100 µl
规格 Spin column
主要样本类型 Fatty tissue samples
处理 Manual
纯化总RNA、miRNA、poly A+ mRNA、DNA或蛋白 RNA
样本量 10–100 mg
技术 Silica technology
每次运行或制备时间 45 minutes
产量 2.4–10 mg

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