IBI Scientific IB47700 DNA/RNA/Protein Extraction Kit (4 Preps)

The DNA RNA Protein Extraction Kit provides an efficient method for purifying genomic DNA, total RNA, and total protein simultaneously from cultured cells, animal tissues, whole blood, and biological fluids. Chaotropic salt is used to lyse cells and inactivate DNases and RNases, allowing DNA to bind to the genomic DNA spin column. The flow-through can then be transferred to the RNA Spin column for RNA binding. Contaminants are effectively removed using wash buffers followed by pure genomic DNA elution in a low salt buffer and pure total RNA elution in RNase-free water.

DNA/RNA purification can be completed in 25 minutes without phenol/chloroform extraction or alcohol precipitation, and protein purification can be completed in 50 minutes. The purified DNA, with approximately 20 - 30 Kb, is suitable for use in PCR or other enzymatic reactions and the purified RNA (including miRNA) is ready for use in RT-PCR, Real-Time PCR, northern blotting, primer extension, mRNA selection, and cDNA synthesis. The purified proteins can be directly analyzed on a SDS-PAGE and subsequent western blot.

The quality of the DNA RNA Protein Extraction Kit is tested on a lot-to-lot basis by isolating genomic DNA and total RNA from 1 x 106 cultured animal cells. The purified DNA and total RNA is quantified with a spectrophotometer and analyzed by electrophoresis on a 1% agarose gel. The purified protein is quantified by Bradford assay analyzed on SDS-PAGE.

The cell RNA can be extracted from hydrogel using the IBI Isolate DNA/RNA Reagent Kit.

• Collect 5-10 hydrogels in a tube and wash the hydrogels with PBS.
• Add 1 ml of  IBI Isolate DNA/RNA Reagent to the hydrogels.
• Incubate the sample on ice for 20-30 minutes.
• Disrupt the hydrogels by passing through a 19 Gauge needle for 10 times.
• Continue to disrupt the hydrogels by passing through a 21 Gauge needle for 10 times, then vortex for 30 seconds.
• Centrifuge the sample at 10,000 x g for 2 minutes at 4°C.
• Transfer the supernatant to a clean 1.5 ml tube.
• Add 200 μl of chloroform to the sample per 1 ml of IBI Isolate DNA/RNA Reagent used in sample homogenization.
• Proceed with the following steps to extract the RNA.

Features:

• Sample Size: up to 5x10⁶ cultured animal cells/up to 25 mg animal tissue /up to 500 µl whole blood/up to 200 µl biological fluids (serum, plasma)
• Expectant Yield: up to 9 µg of genomic DNA/20 µg of total RNA/120 µg of protein from 1x10⁶ of He La cells
• Operation Time: DNA/RNA purification within 25 minutes, protein precipitation within 50 minutes
• Elution Volume: 50 - 200 µl (genomic DNA)/25 - 50 µl (total RNA)
• Format: genomic DNA spin column and total RNA spin column

This kit has 4 reactions.

For more information, please see Protocol