Purification of Oligonucleotides
1.0 mg load
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Resin: |
TOYOPEARL GigaCap Q-650S
|
Column size: |
6.6 mm ID × 18.5 cm (6.3 mL) |
Mobile phase: |
Buffer A: 20 mmol/L NaOH
Buffer B: 20 mmol/L NaOH, 3.0 mol/L NaCl
|
Gradient: |
50% B (2 CV)
50 - 100% B (15 CV)
100% B (2 CV) |
Flow rate: |
200 cm/hr (1.14 mL/min) |
Detection: |
UV @ 254 nm |
Sample: |
crude phosphorothioate deoxyoligonucleotide |
Sample load: |
1.0 mg |
80% DBC elution peak with fraction purity histogram
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|
|
Purification of oligonucleotide at 80% DBC
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Resin: |
TOYOPEARL GigaCap Q-650S
|
Column size: |
6.6 mm ID × 18 cm (6.16 mL) |
Mobile phase: |
Buffer A: 20 mmol/L NaOH
Buffer B: 20 mmol/L NaOH, 3.0 mol/L NaCl
|
Gradient: |
step to 20% B (2 CV)
20 - 100% B (20 CV)
100% B (2 CV) |
Flow rate: |
200 cm/hr (1.14 mL/min) |
Detection: |
UV @ 254 nm |
Sample: |
crude phosphorothioate deoxyoligonucleotide |
Sample load: |
181.4 mg |
Oligonucleotide purity and yield from 80% DBC purification
Resin: |
crude oligo purity |
final oligo purity |
% yield |
TOYOPEARL GigaCap
Q-650S |
66.5% |
96.9% |
81.3% |
|
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