Simultaneous extraction of Genomic DNA and Total RNA from cultured cells, animal tissues, whole blood and biological fluids.
- Sample Size: up to 5×106 cultured animal cells/up to 25mg animal tissue /up to 500µl whole blood/up to 200µl biological fluids (serum, plasma)
- Expectant Yield: up to 9µg of genomic DNA/20µg of total RNA/120µg of protein from 1×106 of HeLa cells
- Operation Time: DNA/RNA purification within 25 minutes, protein precipitation within 50 minutes
- Elution Volume: 50 – 200µl (genomic DNA)/25 – 50µl (total RNA)
- Format: genomic DNA spin column and total RNA spin column
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 – 30Kb, 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 Extraction Kit is tested on a lot-to-lot basis by isolating genomic DNA and total RNA from 1×106 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.