Column for DNA/RNA separation and purification "Presep® DNA/RNA"
Presep® DNA/RNA columns are solid-phase extraction columns suitable for the separation and purification of nucleic acids. Type A columns are mini-columns filled with a silica-based separating agent in a barrel-shaped syringe, which can be used for solid-phase extraction in reversed-phase systems.
Features
- Achieves a high sample load
- Enables a sample load 3 to 5 times higher than commercially available pretreatment columns
- Excellent deprotection efficiency
- Enables purification with a high level of purity and high recovery rate
Product Appearance
Specification
| No. | Product Name | Packing Materials | Syringe Size | Purification amount |
|---|---|---|---|---|
| 1. | Presep® DNA/RNA Type A (85 mg / 1 mL) | Silica gel | 5.5 φ × 57 mm | 0.2-0.5 μmol |
| 2. | Presep® DNA/RNA Type A (255 mg / 3 mL) | Silica gel | 9.0 φ × 63 mm | 1-1.5 μmol |
| 3. | Presep® DNA/RNA Type A (1.0 g / 15 mL) | Silica gel | 15 φ × 87 mm | 4-6 μmol |
| 4. | Presep® DNA/RNA Type A (1.7 mg / 25 mL) | Silica gel | 21 φ × 85 mm | 6-10 μmol |
| 5. | Presep® DNA/RNA Type A (5.1 mg / 70 mL) | Silica gel | 27 φ × 134 mm | 20-30 μmol |
Precautions for Use
Presep® DNA/RNA should be used in the aspiration mode where the exit side of the column is depressurized and aspirated. An aspiration manifold is useful for handling multiple samples.
Solid-phase extraction conditions (recommended)
1. Presep® DNA/RNA Type A (85 mg / 1 mL)
| Steps | Materials | Amount Used |
|---|---|---|
| Conditioning | Acetonitrile | 1 mL |
| 100 mg/mL NaCl aq. | 1 mL × 2 | |
| Sample Load | Sample solution 0.5 mL + 100 mg/mL NaCl aq. 0.5 mL | 1mL |
| Washing 1 | Acetonitrile/100 mg/mL NaCl aq.=5/95(v/v) | 1 mL × 2 |
| Deprotection | 2% DCA*1 aq. (DCA/Water=2/98(v/v)) | 1 mL |
| Washing 2 | RNase Free Water | 1 mL × 2 |
| Elution | 0.5% NH4OH in Acetonitrile/Water=50/50(v/v)*2 | 1 mL |
2. Presep® DNA/RNA Type A (255 mg / 3 mL)
| Steps | Materials | Amount Used |
|---|---|---|
| Conditioning | Acetonitrile | 3 mL |
| 100 mg/mL NaCl aq. | 3 mL × 2 | |
| Sample Load | Sample solution 1 mL + 100 mg/mL NaCl aq. 1 mL | 2 mL |
| Washing 1 | Acetonitrile/100 mg/mL NaCl aq.=5/95(v/v) | 3 mL × 2 |
| Deprotection | 2% DCA*1 aq. (DCA/Water=2/98(v/v)) | 3 mL |
| Washing 2 | RNase Free Water | 3 mL × 2 |
| Elution | 0.5% NH4OH in Acetonitrile/Water=50/50(v/v)*2 | 3 mL |
3. Presep® DNA/RNA Type A (1.0 g / 15 mL)
| Steps | Materials | Amount Used |
|---|---|---|
| Conditioning | Acetonitrile | 12 mL |
| 100 mg/mL NaCl aq. | 12 mL × 2 | |
| Sample Load | Sample solution 5 mL + 100 mg/mL NaCl aq. 5 mL | 10 mL |
| Washing 1 | Acetonitrile/100 mg/mL NaCl aq.=5/95(v/v) | 12 mL × 2 |
| Deprotection | 2% DCA*1 aq. (DCA/Water=2/98(v/v)) | 12 mL |
| Washing 2 | RNase Free Water | 12 mL × 2 |
| Elution | 0.5% NH4OH in Acetonitrile/Water=50/50(v/v)*2 | 12 mL |
4. Presep® DNA/RNA Type A (1.7 g / 25 mL)
| Steps | Materials | Amount Used |
|---|---|---|
| Conditioning | Acetonitrile | 20 mL |
| 100 mg/mL NaCl aq. | 20 mL × 2 | |
| Sample Load | Sample solution 10 mL + 100 mg/mL NaCl aq. 10 mL | 20 mL |
| Washing 1 | Acetonitrile/100 mg/mL NaCl aq.=5/95(v/v) | 20 mL × 2 |
| Deprotection | 2% DCA*1 aq. (DCA/Water=2/98(v/v)) | 20 mL |
| Washing 2 | RNase Free Water | 20 mL × 2 |
| Elution | 0.5% NH4OH in Acetonitrile/Water=50/50(v/v)*2 | 20 mL |
5. Presep® DNA/RNA Type A (5.1 mg / 70 mL)
| Steps | Materials | Amount Used |
|---|---|---|
| Conditioning | Acetonitrile | 60 mL |
| 100 mg/mL NaCl aq. | 60 mL × 2 | |
| Sample Load | Sample solution 20 mL + 100 mg/mL NaCl aq. 20 mL | 40 mL |
| Washing 1 | Acetonitrile/100 mg/mL NaCl aq.=5/95(v/v) | 60 mL × 2 |
| Deprotection | 2% DCA*1 aq. (DCA/Water=2/98(v/v)) | 60 mL |
| Washing 2 | RNase Free Water | 60 mL × 2 |
| Elution | 0.5% NH4OH in Acetonitrile/Water=50/50(v/v)*2 | 60 mL |
*2 Add 50 µL of 28% ammonium hydroxide per 10 mL of 50% acetonitrile in water.
Comparison of separation capacity
1. Recovery of oligonucleotide
Sample of Cartridge Column
| Wako (Presep® DNA/RNA Type A) |
Competitor A | Competitor B | Competitor C | |
|---|---|---|---|---|
| Particle Support | Silica gel | Silica gel | Polymer | Polymer |
Sample of Oligonucleotide
| Sample Name | Length | Linkage Structure |
|---|---|---|
| Sample 1 | 14 mer | PO |
| Sample 2 | 21 mer | PO |
| Sample 3 | 21 mer | PS |
| Sample 4 | 50 mer | PO |
Cartridge Column vs Recovery of oligonucleotide [%]
achieves high recovery.
Wako ≒ B > A > C
- Oligonucleotide samples were purified using the protocol recommended by each company.
- Oligonucleotide concentration was measured with an absorbance meter.
- Recovery was calculated as "oligonucleotide concentration in the eluate/oligonucleotide concentration before loading".
2. Allowable load of oligonucleotide
Sample Load vs Recovery of oligonucleotide [%]
a high support retention capacity.
When the ratio of the load of oligonucleotide to support was increased to 9.7, Wako’s product maintained a recovery rate of 75% or higher in contrast to competitor products (A, B, and C) that had a recovery rate of less than 65%.
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For research use or further manufacturing use only. Not for use in diagnostic procedures.
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