从细胞、组织或酵母中纯化至多1 mg总RNA

  • 数分钟内快速纯化得到高品质总RNA
  • 即用型RNA在各种下游应用中表现良好
  • 中量起始样本,RNA产量稳定
  • 无需酚/氯仿抽提
  • 无需CsCl梯度离心,无需LiCl或乙醇沉淀

RNeasy Midi Kit可从细胞、组织或酵母中快速纯化高品质RNA,使用的硅胶膜RNeasy离心柱的结合能力达1 mg RNA。使用RNAlater RNA Stabilization Reagent或Allprotect Tissue Reagent可方便的稳定组织样本,使用TissueRuptor或TissueLyser体系可有效的破碎组织。处理更小或更大的样本,可使用RNeasy Micro Kit(离心柱结合能力为45 µg RNA)、RNeasy Mini Kit(离心柱结合能力为100 µg RNA)或RNeasy Maxi Kit(离心柱结合能力为6 mg)。

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RNeasy Midi Kit (10)
10 RNeasy Midi Spin Columns, Collection Tubes (15 ml), RNase-free Reagents and Buffers
75142
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RNeasy Midi Kit (50)
50 RNeasy Midi Spin Columns, Collection Tubes (15 ml), RNase-free Reagents and Buffers
75144
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The RNeasy Midi Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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RNeasy Midi procedure.|High-quality RNA from a variety of samples.|RNeasy Midi spin column.|
Total RNA purified with the RNeasy Midi Kit is of high quality and is suitable for many downstream applications. Protocols are also included for cleanup of partially purified RNA, in vitro transcripts, and RNA from enzymatic reactions. An additional buffer is required (composition provided) for isolation of cytoplasmic RNA from eukaryotic cells. Lyticase, zymolase, or glass beads (required for yeast samples) are not provided. Amounts of RNA isolated from samples can vary due to the developmental stage, species, and growth conditions of the sample source. Since the RNeasy procedure enriches for RNA species >200 nt, RNA yield does not include 5S rRNA, tRNAs, or other low-molecular-weight RNAs.|Formaldehyde agarose gel and northern blot of total RNA purified with the RNeasy Maxi Kit. Total RNA (10 µg) isolated from each source was loaded per lane. All tissues were from mouse. Yeast: Saccharomyces cerevisiae; E. coli strain: HB101. 32P-labeled probes recognized (G) GAPDH; (E) translation elongation factor EF-1α; and (O) outer membrane protein A sequences. (E and O were kindly provided by P. Philippsen, University of Basel, Switzerland and U. Henning, Max Planck Institute of Biology, Tübingen, Germany, respectively.) B. subtilis was not probed. M: 0.24–9.5 kb RNA ladder. 7.5 kb band (indicated) in embryo, Huh7, and HeLa cell lanes is a nuclear precursor RNA.|The RNeasy Midi spin column contained in the RNeasy Midi Kit.|
Performance

RNeasy Midii Kit纯化得到的高品质总RNA适用于多种下游应用(参见"High-quality RNA from a variety of samples")。RNeasy Midi Kit可简单地从动物或人类细胞、动物或人类组织或酵母中纯化总RNA(参见下表)。

RNeasy Midi Kit纯化得到的总RNA产量
来源 起始材料 平均产量 (µg)
动物细胞
LMH 
HeLa 
COS-7 
淋巴细胞 (未受激)

7 x 107
7 x 107
3 x 107
1 x 108
850
1000
950
50
小鼠组织



200 mg
100 mg
200 mg

700
130
600
酵母细胞
S. cerevisiae

2 x 108

450
 
Principle

RNeasy Midi Kit可从中量起始样本中高效纯化RNA。RNeasy技术整合了苯酚/异硫氰酸胍裂解的严谨性和硅胶膜纯化的速度和纯度,简化了总RNA纯化技术。RNeasy Kits可纯化得到极高品质的RNA,DNA污染达到最低。

Procedure
RNeasy Midi Kit可从5 x 106–1 x 108个动物或人类细胞、20–250 mg动物或人类组织或2 x 107–5 x 108个酵母细胞中简单的纯化总RNA。先对样本进行破碎和匀浆。在裂解物中加入乙醇以提供合适的结合条件。然后将裂解物上样至RNeasy硅胶膜(参见"RNeasy Midi spin column")。 RNA(至多1 mg)结合到膜上,污染物被高效洗去。对于某些对少量DNA十分敏感的RNA应用,可进行方便的柱上DNase处理,去除残留的DNA。之后可用300–500 µl水洗脱得到高浓度的纯化RNA(参见"RNeasy Midi procedure" )。还有一系列特殊应用的实验方案可选。
Applications

使用RNeasy技术纯化得到的RNA的A260/280比为1.9–2.1(10 mM Tris·Cl、pH 7.5的溶液中),适用于多种下游应用,包括:

Northern印迹、点印迹和狭缝印迹
终点式RT-PCR
定量real-time RT-PCR
芯片分析 
Poly A+ RNA筛选
Feature
Specifications
Applications PCR, qPCR, real-time RT-PCR, microarray
Elution volume 300–500 µl
Format Spin column
Main sample type Tissue, cells
Processing Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein RNA
Sample amount 20–250 mg
Technology Silica technology
Time per run or per prep <1 hour
Yield Varies

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试剂盒操作手册
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RNeasy Midi Kit RNeasy Maxi Kit For total RNA isolation from animal cells animal tissues bacteria yeast whole blood and for RNA cleanup RNeasy Protect Midi Kit RNeasy Protect Maxi Kit For stabilization and protection of RNA in tissue For isolation of t
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