RNeasy Mini Kit

从细胞、组织或酵母中纯化至多100 μg的总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
  • 少量起始样本,RNA产量稳定
  • 无需酚/ 氯仿抽提,无需氯化铯梯度离心,无氯化锂或者乙醇沉淀步骤
RNeasy Mini Kit可从细胞、组织或酵母中快速纯化高品质RNA,硅胶膜RNeasy离心柱的结合能力达100 μg 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。可在QIAcube全自动核酸纯化仪上应用RNeasy Mini Kit实现RNA纯化自动化。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 Midi Kit(离心柱结合能力为1 mg RNA)和RNeasy Maxi Kit(离心柱结合能力为6 mg RNA)。
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产品 货号 目录价:
RNeasy Mini Kit (50)
50 RNeasy Mini Spin Columns, Collection Tubes (1.5 ml and 2 ml), RNase-free Reagents and Buffers
74104
询价
RNeasy Mini Kit (250)
250 RNeasy Mini Spin Columns, Collection Tubes (1.5 ml and 2 ml), RNase-free Reagents and Buffers
74106
询价

RNeasy 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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RNeasy Mini procedure.|High-quality RNA from a variety of samples.|RT-PCR of RNA from as few as 100 cells.|RNeasy Mini spin column.|
Total RNA purified with the RNeasy Mini 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. 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.|RT-PCR of total RNA isolated with the RNeasy Mini Kit from the indicated numbers of HeLa cells. 10 µl (1/5) of eluate was digested with RNase-free DNase and reverse transcribed with oligo-dT primer. 2.5 µl (1/20) of the cDNA mix was used in 50 µl PCR. A 452 bp fragment of GAPDH was amplified. C-: negative control; C+: positive control; M: 100 bp ladder.|The RNeasy Mini spin column contained in the RNeasy Mini Kit.|
Performance

RNeasy Mini Kit可从至多30 mg动物或人类组织样本,或从100–1 x 107个动物或人类细胞样本或酵母中纯化总RNA(参见"RT-PCR of RNA from as few as 100 cells"和"High-quality RNA from a variety of samples")。

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

1 x 106
1 x 106
1 x 106
1 x 106

12
15
35
0.5
小鼠组织



10 mg
10 mg
10 mg

40
10
35
真菌细胞
S. cerevisiae

1 x 107

25
 
Principle

RNeasy Mini Kit可从少量的样本中高效纯化总RNA。RNeasy纯化技术整合了苯酚/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。纯化得到高品质的RNA,并且DNA残留水平达到最低。

Procedure
RNeasy Mini Kit可从10–1 x 107个动物或人类细胞中、0.5–30 mg动物或人类组织中或小于5 x 107个酵母细胞中方便的纯化总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 Mini spin column")。RNA结合到膜上(最多可结合100 µg),污染物被高效洗去。对于某些对少量DNA十分敏感的RNA应用,可在RNeasy操作过程中进行柱上DNA酶处理,去除DNA残留。之后可在30–100 µl的纯水洗脱液中得到高浓度的纯化RNA(参见"RNeasy Mini procedure")。还有各种特殊应用的实验方案可供选择。
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
特点
参数
应用 PCR, qPCR, real-time RT-PCR, microarray
洗脱体积 30–100 µl
规格 Spin column
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 0.5–30 mg
技术 Silica technology
每次运行或制备时间 20 minutes
产量 Varies

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试剂盒操作手册
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RNeasy Mini Kit - For purification of total RNA from animal cells, animal tissues, bacteria, and yeast, and for RNA cleanup; RNeasy Protect Mini Kit - For immediate stabilization of RNA in harvested animal tissues and subsequent total RNA purification; RNeasy Plant Mini Kit - For purification of total RNA from plants and filamentous fungi
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补充实验方案
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Up to 1 x 109 bacteria are disrupted and homogenized by bead-milling in a guanidine-thiocyanate-containing lysis buffer. After addition of ethanol, the sample is loaded onto an RNeasy Mini spin column. Total RNA binds to the RNeasy silica-membrane, contaminants are efficiently washed away, and high-quality RNA is eluted in RNase-free water.
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用户开发的实验方案
1
One milliliter of milk contains approximately 50,000-200,000 leukocytes (with an average of approximately 100,000 leukocytes). In a BVDV-infected animal up to 1-30% of the leukocytes may be infected with the virus. Each infected leukocyte typically contains 10-10,000 BVDV entities.
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学术海报
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参考文献
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图片
RNeasy Mini Procedure
RNeasy Mini实验方案。
使用RNeasy Mini Kit纯化的总RNA具有高质量,适用于多种下游操作。实验方案还包括部分纯化的RNA、体外转录本和酶反应RNA的回收。不提供溶壁酶、酵母裂解酶或玻璃珠(酵母样本需要)。因样本来源的发育阶段、种属和生长条件的不同,从样本中分离的RNA量也各异。由于RNA酶步骤富集的RNA种属>200 nt,因此RNA产量不包括5S rRNA、tRNA或其他低分子量RNA。
High-quality RNA from a variety of samples.
从多种样本中纯化高品质RNA。
使用RNeasy Maxi Kit纯化的总RNA的甲醛琼脂糖凝胶电泳和northern印迹。从各种来源的样本中分离总RNA(10 µg)上样至各泳道。所有组织均来源于小鼠。酵母:Saccharomyces cerevisiaeE. coli菌株:HB101。32P标记的探针识别的(G)GAPDH;(E)翻译延长因子EF-1α;和(O)外膜蛋白A序列。(E和O分别由瑞士巴塞尔大学的P. Philippsen和德国蒂宾根马克斯-普朗克生物学研究所的U. Henning提供。)B. subtilis未采用探针检测。M:0.24–9.5 kb RNA分子量标准。胚胎、Huh7和HeLa细胞泳道中的7.5 kb条带(图示)为核前体RNA。
RT-PCR of RNA from >100 cells.
对少至100个细胞的RNA进行RT-PCR。
使用RNeasy Mini Kit从图示数目的HeLa细胞中分离的总RNA的RT-PCR。使用不含RNA酶的DNA酶酶切10 µl(1/5)洗脱液,并使用oligo-dT引物进行逆转录。使用2.5 µl(1/20)的cDNA混合物进行50 µl PCR。扩增452 bp的GAPDH片段。C-:阴性对照;C+:阳性对照;M:100 bp分子量标准。
RNeasy Mini spin column
RNeasy Mini离心柱。
RNeasy Mini Kit中提供RNeasy Mini离心柱。