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miRNeasy Mini Kit

从多种动物组织和细胞中纯化microRNA和总RNA

  • 有效纯化miRNA和总RNA
  • 高效富集miRNA和< 200个核苷酸的RNA
  • 高纯度RNA适合多种下游应用
  • 灵活的操作方法可用于共纯化或单独纯化各组分
miRNeasy Mini Kit可从多种类型的动物组织和细胞,包括难裂解的组织中纯化18个核苷酸(nt)以上的总RNA,也可单独纯化富集后的miRNA组分和总RNA(>200 nt)组分(单独纯化需要RNeasy MinElute Cleanup Kit)。miRNeasy Mini Kit利用离心柱进行低通量RNA纯化。miRNeasy Mini Kit可在QIAcube全自动核酸纯化议上自动完成miRNA与总RNA的共纯化。
Cat No./ID: 217004
miRNeasy Mini Kit (50)
For 50 preps: 50 RNeasy Mini Spin Columns, Collection Tubes (1.5 ml and 2 ml), QIAzol Lysis Reagent, RNase-Free Reagents and Buffers

miRNeasy 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。


0
Effective purification from a range of starting amounts.
Total RNA was purified from [A] 102–107 Jurkat cells using the miRNeasy 96 Kit or [B] a dilution series of rat lung tissue homogenate from 20 mg to 200 ng using the miRNeasy Mini Kit. miRNA-enriched fractions (<200 nt) were also isolated from the same samples. Purified RNA was used as a template in quantitative, real-time RT-PCR assays for the miRNA miR-16.
1
Efficient copurification from a wide range of tissues.

Total RNA including miRNA was purified from 25 mg of a range of RNAlater stabilized rat tissues using the miRNeasy 96 Kit. Purified RNA was used as a template in quantitative, real-time RT-PCR assays for the miRNA miR-16 and for the larger mRNA of the PGK1 gene. Results showed successful detection of both PGK1 mRNA (large RNA) and miR-16 (small RNA) from the same eluates.

2
miRNeasy Mini Kit outperforms TRIzol.
Total RNA including miRNA was purified from a range of amounts of Jurkat cells using the miRNeasy Mini Kit or TRIzol Reagent. Purified RNA was used as a template in quantitative, real-time RT-PCR assays for the miRNA miR-16. Results showed that CT values were lower after purification using the miRNeasy Kit, indicating that higher amounts of miRNA were purified than using TRIzol. miRNA was effectively purified from as little as 1 x 102 cells using the miRNeasy Kit. In contrast, no miRNA was detected after 40 PCR cycles from 1 x 102 cells when TRIzol was used for purification.
3
Effective enrichment of small RNA.
miRNA-enriched fractions were purified from 3 x 106 HeLa cells using either the miRNeasy Mini Kit or a kit from Supplier AIV. Equal RNA amounts from miRNA-enriched fractions were run on a 1.5% gel. For comparison, 20mer and 21mer synthetic RNAs were also run on the gel. The miRNA-enriched fraction prepared using the miRNeasy Kit showed a smear of RNAs of different sizes down to approximately 18 nt. In contrast, in the miRNA-enriched fraction prepared using the kit from Supplier AIV, primarily tRNA and 5S rRNA were visible.
4
Highly pure RNA without phenol carryover.
Total RNA including miRNA was purified from 1 x 106 Jurkat cells using the miRNeasy Mini Kit or TRIzol Reagent. The absorbance spectra showed the OD maximum for RNA purified using the miRNeasy Mini Kit was at 260 nm. In contrast, the OD maximum was greater than 260 nm when TRIzol was used for purification, indicating phenol carryover. In addition, the OD230 measurement was higher in the TRIzol-prepared RNA, indicating salt carryover from the TRIzol Reagent.
5
miRNeasy procedures.

The miRNeasy Mini Kit enables purification of total RNA which includes RNA from 18 nucleotides (nt) upwards. Alternatively, an miRNA-enriched fraction and a total RNA (>200 nt) fraction can be can be purified separately. A separate miRNA-enriched fraction may be required to reduce background in particularly sensitive downstream applications. Separate purification procedures using the miRNeasy Mini Kit require additionally the RNeasy MinElute Cleanup Kit. For purification in a 96-well format, we recommend the miRNeasy 96 Kit.

Performance

使用miRNeasy Mini Kit纯化含有miRNA的总RNA,可用于与管家基因或其他目的mRNA进行miRNA表达水平的直接比较。对于某些比较灵敏的下游应用,还需要富集得到不含有大分子RNA的miRNA组分。miRNeasy Mini Kit可高效富集至少含18个核苷酸以上的RNA。相比于使用其他试剂盒纯化的样本,使用miRNeasy Mini Kit纯化后的小RNA更清晰可见(参见"Effective enrichment of small RNA")。miRNeasy Kits可从组织和细胞中纯化RNA,即使初始样本量较低也可确保高效纯化。随着纯化RNA的细胞或组织量的降低,CT值呈线性增长,说明可从较大范围的初始样本中高效纯化miRNA(参见"Effective purification from a range of starting amounts")。

该试剂盒可从多种组织和细胞中纯化RNA,包括难裂解的组织(参见"Efficient copurification from a wide range of tissues")。miRNeasy流程可有效避免盐类或苯酚污染等影响后续分析的可能性(参见"Highly pure RNA without phenol carryover")。miRNeasy Kits与其他纯化miRNA的方法相比,如TRIzol Reagent,不仅纯度更高,还可得到更高产量(参见"miRNeasy Kit outperforms TRIzol")。使用miRNeasy Kits制备的RNA纯度高,可直接用于各种灵敏的下游应用。

Principle

miRNeasy Mini Kit将苯酚异硫氰酸胍样本裂解技术与硅胶膜总RNA纯化技术相结合。该试剂盒包含的QIAzol Lysis Reagent,为苯酚与异硫氰酸胍的单相溶液,用于促进组织的裂解并抑制RNases,同时除去有机溶剂萃取产物中的绝大部分细胞DNA以及蛋白质。

Procedure
miRNeasy Mini Kit可纯化18个核苷酸(nt)以上的总RNA,也可分别纯化富集后的miRNA组分和总RNA(>200 nt)组分(参见"miRNeasy procedures")。单独纯化的富集后的miRNA组分可能需要减弱背景干扰,以便用于某些比较灵敏的下游应用。在这种特定情况下,对miRNA和RNA(<200 nt)进行富集需要RNeasy MinElute Cleanup Kit。 miRNeasy Mini Kit可在QIAcube全自动核酸纯化仪上自动纯化含有miRNA的总RNA。独创的QIAcube全自动核酸纯化仪应用先进技术处理QIAGEN离心柱,实现低通量样本自动化制备与实验室流程的无缝结合。使用QIAcube全自动核酸纯化仪制备样本与手工操作步骤相同(即裂解、结合、洗涤和洗脱),确保使用miRNeasy Mini Kit能持续纯化高品质的RNA。更多关于自动化流程的信息,参见相关的实验方案。
Applications

miRNeasy Mini Kit对含有miRNA的总RNA的纯化适合多种下游应用,包括:

  • Northern印迹分析
  • 定量real-time RT-PCR
  • 微阵列分析
Features
Specifications
应用 PCR, qPCR, real-time RT-PCR, microarray
洗脱体积 30 µl
规格 Spin column
主要样本类型 Tissue, cells
处理 Manual (centrifugation or vacuum)
纯化总RNA、miRNA、poly A+ mRNA、DNA或蛋白 miRNA, total RNA
样本量 50–100 mg
技术 Silica technology
产量 Varies

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试剂盒操作手册 (1)
For purification of total RNA, including miRNA, from animal and human cells and tissues
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