RNeasy Plus Micro Kit (50)，74034，Qiagen，凯杰

RNeasy Plus Kits

For fast purification total RNA from cells and tissues using gDNA Eliminator columns or plates

Features

Efficient gDNA removal with unique gDNA Eliminator columns or plates (no need for DNase)

High-quality total RNA in minutes using fast and simple extraction protocols

Phenol-free RNA isolation

High-throughput processing in 96-well format

Ideal for sensitive applications such as real-time RT-PCR and RNA-seq

RNeasy Plus Kits are next generation QIAGEN RNA extraction kits, combining high-quality RNA purification with effective gDNA removal with unique gDNA Eliminator columns or plates. Total RNA of high purity and yield is obtained from a wide range of cultured cells and easy-to-lyse tissues. The RNeasy Plus Micro Kit is specially designed for limited amounts of samples and isolates up to 45 µg pure total RNA The RNeasy Plus Mini Kit purifies up to 100 µg total RNA (>200 nt) from a single extraction with efficient gDNA removal. Both kits can be automated on the QIAcube Connect and samples can be conveniently stabilized using RNAprotect Tissue Reagent or Allprotect Tissue Reagent and efficiently disrupted using a TissueRuptor II or TissueLyser II or LT system (available separately). The RNeasy Plus 96 Kit is for high-throughput purification of total RNA from cultured cells grown in 96-well plates. For optimal results it is recommended to use this product together with the QIAvac 96 System.

Request a quote for a trial RNeasy Plus Mini Kit.

RNA extraction from cells and tissues with RNeasy Plus Micro and Mini Kits allows high, reproducible RNA yields and efficient genomic DNA elimination for sensitive applications (see figures “ Effective cell genomic DNA removal”, “ Effective tissue genomic DNA removal”, " High, reproducible RNA yields", and “ Array-ready RNA”). Total RNA with Agilent RIN values of close to 10 is routinely obtained from tissues and cultured cells.

RNeasy Plus Micro and Mini Kits also provide an enrichment for mRNA since most RNAs <200 nucleotides (such as rRNAs and tRNAs) are excluded. For isolation of total RNA containing small RNAs (e.g., miRNA) from cultured cells, additional RNA isolation protocols are available and described in the RNeasy Plus Micro Handbook and as a separate Supplementary Protocol to the RNeasy Plus Mini Kit.

The RNeasy Plus 96 Kit is compatible with a wide range of cell lines, providing purification of high-quality RNA with Agilent RIN values of close to 10 (see figure " High-quality RNA"). Complete genomic DNA removal by the kit (see figure "Effective genomic DNA removal") enables specific transcript detection in highly sensitive applications, such as quantitative, real-time RT-PCR analysis of low-abundance targets. In addition, reliable RNA purification from 102 to 106 cells allows real-time RT-PCR quantification over a wide dynamic range (see figure " Reliable RNA purification").

Total RNA extraction: effective cell genomic DNA removal.

Total RNA was purified from Jurkat cell samples (1 x 106 cells per sample) using [A] the RNeasy Plus Mini Kit, or [B] an RNA purification kit with integrated genomic DNA removal from Supplier AV. Duplicate real-time RT-PCR assays for β-actin were performed with (+RT) or without (-RT) reverse transcriptase. The -RT curves demonstrate that RNA purified using the RNeasy Plus Mini Kit was virtually free of genomic DNA.

Total RNA extraction: effective tissue genomic DNA removal.

Total RNA was purified in duplicate from various mouse tissues (10 mg per sample) using the RNeasy Plus Mini Kit or kits from other suppliers. Real-time PCR assays for c-jun were performed to determine the amount of DNA contamination in the purified RNA.

High, reproducible RNA yields.

Total RNA was purified in duplicate from different amounts of Jurkat cells using the RNeasy Plus Mini Kit or a similar kit from Supplier AV. Real-time RT-PCR assays for β-actin were performed (40 cycles). The lower CT values with the RNeasy Kit demonstrate greater RNA yields. With the kit from Supplier AV, no transcript was detectable in RNA purified from 102 cells.

Array-ready RNA.

Total RNA was purified in duplicate from 1 x 106 HeLa cells using the RNeasy Plus Mini Kit. cRNA was prepared from the duplicate RNA samples (3.5 μg each) using the GeneChip IVT Labeling Kit. The cRNA samples (15 μg each) were analyzed on GeneChip Human Genome U133A probe arrays. The scatter plot shows the correlation between the two samples (Pearson correlation coefficient [r] is 0.996). Red: gene present in both samples; Blue: gene absent or marginal in one sample; Yellow: gene absent or marginal in both samples.

High quality RNA.

Total RNA was purified from the indicated cell lines using the RNeasy Plus 96 Kit and analyzed on the Agilent 2100 Bioanalyzer. The RIN values were close to 10, indicating the high integrity of the purified RNA.

Effective genomic DNA removal.

Total RNA was purified from Jurkat cells (105, 104, 103, and 102) using the RNeasy Plus 96 Kit. Real-time RT-PCR analysis of the β-actin transcript was performed using a primer–probe set that could amplify and detect both mRNA and genomic DNA sequences. The flat amplification plot for the no RT control (–RT) indicates the absence of genomic DNA.

Reliable RNA purification.

Total RNA was purified from 10-fold serial dilutions of Jurkat cells (106 to 102 cells) using the RNeasy Plus 96 Kit. Real-time RT-PCR analysis of the β-actin transcript was performed. The linear graph of CT values against cell dilution indicates reliable RNA purification over a wide range of amounts of starting material (R2 = 0.9985).

RNA purified using RNeasy Plus Kits is ideally suited for downstream applications that are sensitive to low amounts of DNA contamination, such as RNA-seq and quantitative real-time RT-PCR. The purified RNA can also be used in other applications.

The RNeasy Plus Micro Kit is suitable for small samples, including:

Laser-microdissected cryosections

Fine-needle aspirates

The ease with which RNA is purified in 96-well format makes the RNeasy Plus 96 Kit well suited for high-throughput gene expression analysis in areas such as:

Drug discovery

Biomedical research

Comparison of RNeasy Plus Kits