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. Breakthrough Curve
Sample Load vs Recovery of oligonucleotide [%]
Our product is engineered with a carrier pore size optimized for medium-sized molecules, enabling them to retain oligonucleotides 2-5 times more efficiently than products from other manufacturers. This enhanced retention significantly reduces the number of purification cycles required as well as the volume of solvent used.
3. DMT Group Deprotection Efficiency
FUJIFILM Wako > C
Our product is engineered with a carrier pore size optimized for medium-sized molecules, enabling them to retain oligonucleotides 2-5 times more efficiently than products from other manufacturers. This enhanced retention significantly reduces the number of purification cycles required as well as the volume of solvent used.Comparative testing against product C from another company confirmed that no DMT-ON impurities remain in our product. This is because the non-swelling nature of the silica gel enhances the efficiency of the DMT group deprotection reaction.
Optimal Extraction Conditions
Optimal Extraction Conditions
Conditions with Ease!
The optimal acetonitrile concentration for the wash step can be estimated from the HPLC retention time.
HPLC Condition
- Column size
- Wakopak® Ultra C18-2 2.1 φ x 100 mm
- Mobile phase
- A) 100 mM TEAA aq.
B) Acetonitrile - Gradient
- 0-15 min B=5-40%, 15-20 min B=100%,20-25 min B=5%
- Flow rate
- 0.3 mL/min
- Temperature
- 40℃
- Injection vol.
- 0.5 μL
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For research use or further manufacturing use only. Not for use in diagnostic procedures.
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