DNA/RNA/Protein Extraction Kit (DRP050, DRP100)

The Presto™ DNA RNA Protein Extraction Kit provides an efficient method for purifying genomic DNA, total RNA and total protein simultaneously from cultured cells, animal tissue, whole blood and biological fluids such as serum and plasma. Chaotropic salt is used to lyse cells and degrade protein, allowing DNA to bind to the genomic DNA mini spin column. The flow-through containing RNA can then be transferred to the RNA mini spin column for RNA binding. The proteins in the flow-through can then be precipitated using acetone. Contaminants are removed using a wash buffer and the purified genomic DNA is eluted in a low salt Elution Buffer and the purified total RNA is eluted in RNase-free Water. The entire procedure can be completed in 25 minutes without phenol/chloroform extraction or alcohol precipitation. The purified DNA, 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. 

Advantages (Cat. # DRP050, DRP100)

• Purify genomic DNA, total RNA within 25 minutes! Protein precipitation within 50 minutes!
• Variety of Samples: up to 500 μl of whole blood, up to 25 mg of tissue, rodent tails, ear punches, up to 5 x 106 cultured cells, up to 200 μl of biological fluids such as serum and plasma
• High Yield: up to 9 μg of genomic DNA, up to 20 μg of total RNA from 1.5 x 106 of cultured HeLa cells, up to 1 μg of genomic DNA, up to 10 μg of total RNA from 500 µl human blood samples
• High Quality DNA: A260/A280 = 1.9-2.0
• Genomic DNA glass fiber spin columns and total RNA glass fiber spin colums certified DNase and RNase-free
• Elution Volume: 50-200 μl (genomic DNA) and 25-50 μl (total RNA)
• Storage: dry at room temperature (15-25°C)

Applications

The purified DNA, 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.

Components

• RBC Lysis Buffer
• DR Buffer
• RW1 Buffer
• RPE Buffer
• W1 Buffer
• Wash Buffer
• RNase-free Water
• Elution Buffer (10 mM Tris-HCl, pH8.5 at 25ºC)
• DV Buffer (8M Urea)
• Protein Loading Dye
• GD Columns
• RB Columns
• 2 ml Collection Tubes

Quality Control

The quality of the Presto™ DNA/RNA/Protein Extraction Kit is tested on a lot-to-lot basis according to Geneaid's ISO-certified quality management system by isolating genomic DNA, total RNA and total protein from 1.5 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 and analyzed on SDS-PAGE.

	Figure 1. Genomic DNA and Total RNA from 1.5 x 106 HeLa cells was extracted using the Presto™ DNA/RNA/Protein Extraction Kit. 10 µl aliquots from 200 µl eluates of purified genomic DNA and 10 µl  aliquots from 50 µl eluates of purified total RNA was analyzed by electrophoresis on a 1% agarose gel. 1-2 = DNA from 1.5 x 106 HeLa cells 3-4 = RNA from 1.5 x 106 HeLa cells M = Geneaid 1 Kb DNA Ladder Sample µg/ml 260/280 260/280 Yield 1. DNA 42.7 1.88 2.19 8.54 µg 2. DNA 46.7 1.88 2.18 9.34 µg 3. RNA 417.5 2.09 2.03 20.9 µg 4. RNA 429.9 2.09 2.04 21.5 µg
	Figure 2. DNA and RNA from 3 human blood samples (500 µl) was extracted using the Presto™ DNA/RNA/Protein Extraction Kit. 10 µl aliquots from a 100 µl eluate of purified genomic DNA and a 50 µl eluate of purified total RNA were analyzed by electrophoresis on a 1% agarose gel. M = Geneaid 1 Kb DNA Ladder Sample µg/ml 260/280 260/280 Yield 1. DNA 13.1 1.84 1.91 0.7 µg 2. DNA 20.8 1.93 1.97 1.0 µg 3. DNA 12.1 1.89 1.89 0.6 µg 4. RNA 93.4 1.83 2.18 9.3 µg 5. RNA 103.1 1.85 2.12 10.3 µg 6. RNA 89.1 1.83 2.16 8.9 µg
	Figure 3. Protein was extracted from 1.5 x 106 HeLa cells in 200 µl of DV buffer using the Presto™ DNA/RNA/Protein Extraction Kit. 20 µl aliquots from a 200 µl eluate of purified protein were analyzed on a NuPAGE4-12% Bis-Tris gel and stained with coomassie blue (120 µg of total protein with a concentration of 0.6 µg/µl).