High Pure RNA Isolation Kit

High Pure RNA Isolation Kit

The High Pure RNA Isolation Kit is expertly engineered for the efficient purification of total RNA, particularly from cultured cells, while also accommodating other sample materials such as blood, yeast, and bacteria with specific pre-lysis treatments detailed in the protocol. This advanced kit integrates a DNase digestion step to significantly reduce contamination from residual genomic DNA, ensuring the isolated RNA’s integrity. Ideal for a range of analytical techniques, including RNase protection, northern blotting, and primer extension, this kit enables the processing of up to 24 samples concurrently within approximately 1 hour. This streamlined process markedly diminishes the time and effort required compared to traditional methods that necessitate the use of organic extraction solutions, RNA precipitation, or ultracentrifugation. The High Pure RNA Isolation Kit stands out for its ability to deliver purified RNA suitable for crucial applications in RNA isolation, reinforcing its utility in studies requiring high-quality RNA samples.

Benefits of the High Pure RNA Isolation Kit

• Quickly process multiple samples. A single sample preparation is completed in 25 minutes, and multiple samples can be prepared in 45 minutes.
• Prevent RNA loss. Precipitation or solvent extraction steps are not required.
• Avoid DNA contamination. Integrated DNA digestion and DNase removal eliminates interference from genomic DNA.
• Obtain concentrated RNA. RNA is eluted in a 50 μl volume.

How the High Pure RNA Isolation Kit works:

Isolation of RNA is a prerequisite for the analysis of gene expression. Frequently, applied techniques, such as reverse transcriptase-PCR (RT-PCR), northern blotting, RNase protection, and primer extension require the use of intact, undegraded RNA from different sample materials, such as cultured cells, blood, yeast, and bacteria. Samples are lysed and homogenized in the presence of chaotropic salts, then applied to the spin filter tube. Nucleic acids bind specifically to the surface of the filter. Co-purified DNA is ultimately digested with DNase I. The bound RNA is purified from salts, proteins, digested DNA, and other impurities by washing steps, followed by an elution.

1. Cultured cells are lysed by a special Lysis/Binding Buffer. At the same time, RNases are inactivated. Other sample materials require a specific pre-lysis treatment.
2. Nucleic acids are bound to the glass fibers pre-packed in the High Pure Filter Tube.
3. Residual contaminating DNA is digested by DNase I and applied directly onto the glass fiber fleece.
4. Bound nucleic acids are washed with a special Wash Buffer to get rid of RT-PCR inhibitory contaminants.
5. Further washing of bound nucleic acids purifies them from salts, proteins, and other cellular impurities.
6. RNA is recovered using the Elution Buffer.

Table: Fresh human blood was collected from two donors with WBC = 10.7 × 10 3 cells/μL and WBC = 4.8 × 10 3 cells/μL. Each isolation was performed with 12 replicates followed by a duplicated analysis on the LightCycler ® 480 Instrument II using the LightMix ® Kit human glucose-6-phosphate dehydrogenase (G6PDH). Isolation was performed according to the Instructions for Use of the respective kit followed by quantitative analysis of G6PDH on the LightCycler ® 480 Instrument I