miRNeasy Mini Kit (50)，217004，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.

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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.

PCR analysis of DNA purified from FFPE tissue samples.

Genomic DNA was purified from different FFPE tissue samples using the QIAamp DNA FFPE Tissue Kit. Products of the prnp gene (78 bp, 464 bp, and 727 bp) were amplified by real-time quantitative PCR.

QIAamp DNA FFPE Tissue Kit procedure.

Paraffin is dissolved in xylene and removed. The sample is lysed under denaturing conditions with a short proteinase K digestion. Incubation at 90°C reverses formalin crosslinking. DNA binds to the membrane and contaminants flow through. Residual contaminants are washed away. DNA is eluted in Buffer ATE or water, and is immediately ready for use in amplification reactions or for storage at –20°C. Purified DNA is free of proteins, nucleases, and other impurities.

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