miRNeasy Micro Kit (50)，217084，Qiagen，凯杰

miRNeasy Kits

For purification of miRNA and total RNA from tissues and cells

Features

Effective microRNA (miRNA) and total RNA purification, even from small samples

Efficient enrichment of miRNA and RNAs<200 nucleotides

Protocols for copurification or isolation of separate fractions

High-purity RNA suitable for all downstream applications

Automatable protocols and high-throughput processing in 96-well format

miRNeasy Kits enable purification of total RNA from all types of animal tissues and cells, including difficult-to-lyse tissues and small tissue and cell samples. Alternatively, an miRNA-enriched fraction and a total RNA (>200 nt) fraction can be purified separately (for separate purification, an RNeasy MinElute Cleanup Kit is required when using the miRNeasy Micro or Mini Kit and an additional RNeasy 96 plate when using the miRNeasy 96 Kit). Purification of 1–12 samples can be automated on the QIAcube Connect. High-throughput processing in convenient 96-well format can be performed using centrifugation or vacuum procedures.

The NEW miRNeasy Tissue/Cells Advanced Mini Kit – get excellent results faster without phenol/chloroform using our QIAzol free Advanced chemistry. Try it today.

QIAzol based microRNA purification (miRNA purification) using miRNeasy Kits efficiently purifies RNA from small tissue (up to 5 mg tissue) and cell (up to 1 x 106 cells) samples (see figure " High, reproducible recovery of miRNA from small cell numbers"). RNA can be purified from a variety of tissues and cells, including difficult-to-lyse tissues.

Purification of total RNA including miRNA allows direct comparison of miRNA expression levels with those of housekeeping reference genes or any other mRNA of interest. Alternatively, for some sensitive downstream applications, an miRNA-enriched fraction without larger RNA may be required. miRNeasy Kits enable efficient enrichment of RNA down to approximately 18 nt in size. The presence of very small RNAs is clearly visible after purification using an miRNeasy Kit, in contrast to samples prepared using an alternative kit (see figure " Effective enrichment of small RNA"). miRNeasy Kits efficiently purify RNA from both tissues and cells, even when low amounts of starting material are used (see figure " Effective purification from a range of starting amounts").

RNA can be purified with miRNeasy Kits from a variety of tissues and cells, including difficult-to-lyse tissues (see figure " Efficient copurification from a wide range of tissues"). miRNeasy procedures eliminate the possibility of contamination with salts or phenol which could interfere with later analyses (see figure " Highly pure RNA without phenol carryover"). In addition to higher purity, miRNeasy Kits offer superior yields to alternative methods of miRNA purification, such as using TRIzol Reagent (see figure " miRNeasy Mini Kit outperforms TRIzol").

RNA prepared using miRNeasy Kits is highly pure and ready for use in sensitive downstream applications.

High, reproducible recovery of miRNA from small cell numbers.

Total RNA was purified from 10-fold serial dilutions of Jurkat cells (106 to 10 cells) using the miRNeasy Micro Kit. RNA was purified from 2 replicates of each dilution, either [A] manually or [B] using the QIAcube. Purified RNA was used to detect miR-16 using the miScript PCR System on a Rotor-Gene Q cycler. miR-16 was detected in as few as 10 cells and CT values were consistently reproducible. No signal was detected in negative controls (blue and yellow curves).

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.

Effective purification from a range of starting amounts.

Efficient copurification from a wide range of tissues.

Total RNA including miRNA was purified from 25 mg of a range of RNAprotect Tissue Reagent 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.

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.

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.

miRNeasy procedures.

miRNeasy Kits allow purification of miRNA with total RNA for use in a variety of applications, including:

RNA-seq

Quantitative, real-time RT-PCR (e.g., using the miScript PCR System)

Northern blot analysis

Microarray analysis

Comparison of miRNeasy Kits