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产品中心 >> 试剂 >> 分子生物学试剂 >> qPCR kits & 核酸抽提
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RNeasy Mini Kit（50）
分类：qPCR kits & 核酸抽提
品牌：Qiagen
货号：74104
原价：￥4600
现价：￥3536.00 节省：￥1064
规格：50Tests
产品参数
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）。
For purification of up to 100 µg total RNA from cells, tissues, and yeast
Performance
Total RNA is easily purified with the RNeasy Mini Kit from up to 30 mg animal or human tissues, from 100 to 1 x 107animal or human cells, or yeast (see table; and figures RT-PCR of RNA from as few as 100 cells and High-quality RNA from a variety of samples).
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Animal cells
LMH
HeLa
COS-7
Lymphocytes (unstimulated)
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1 x 106
1 x 106
1 x 106
1 x 106 |
12
15
35
0.5
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Mouse tissue
Liver
Lung
Spleen
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10 mg
10 mg
10 mg |
40
10
35 |
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Yeast cells
S. cerevisiae
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1 x 107 |
25 |
Principle
The RNeasy Mini Kit allows efficient purification of total RNA from small amounts of starting material. RNeasy technology simplifies total RNA isolation by combining the stringency of guanidine-isothiocyanate lysis with the speed and purity of silica-membrane purification. RNeasy Kits provide the highest-quality RNA with minimum copurification of DNA.
Procedure
With the RNeasy Mini Kit, total RNA is easily purified from 10 to 1 x 10 7 animal or human cells, 0.5–30 mg animal or human tissues, and <5 x 10 7 yeast cells. Samples are first lysed and then homogenized. Ethanol is added to the lysate to provide ideal binding conditions. The lysate is then loaded onto the RNeasy silica membrane (see figure RNeasy Mini spin column). RNA binds (up to 100 µg capacity), and all contaminants are efficiently washed away. For certain RNA applications that are sensitive to very small amounts of DNA, the residual amounts of DNA remaining can be removed using a convenient on-column DNase treatment. Pure, concentrated RNA is eluted in 30–100 µl water (see figure RNeasy Mini procedure). A variety of special application protocols is also available.
Images
RNeasy Mini procedure.
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.
High-quality RNA from a variety of samples.
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 RNA from as few as 100 cells.
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.
RNeasy Mini spin column.
The RNeasy Mini spin column contained in the RNeasy Mini Kit.
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