Oligo Synthesis FAQs
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Custom DNA & RNA Oligos: Synthesis of custom-length oligos (5-150 bases) at 25 – 1000 nmol scales with a wide range of modifications available with HPLC, PAGE, and desalted purification options. Turnaround time starts at 4 business days, depending on scale and modification requirements.
Oligos Retained for Sanger: Order oligos for your GENEWIZ Sanger sequencing order. GENEWIZ will synthesize, dilute, and store the oligo on-site for up to 1 year.
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The GENEWIZ Custom DNA & RNA Oligos service can synthesize oligos between 5-150 bases.
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Oligos are chemically synthesized using the industry-standard, solid-phase phosphoramidite method. Oligos are synthesized through a series of coupling reactions in the 3′ and 5′ direction.
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Yes, GENEWIZ offers synthesis of oligos containing degenerate bases using the standard IUPAC base nomenclature.
| IUPAC Code | Base |
|---|---|
| R | A or G |
| Y | C or T |
| M | A or C |
| K | G or T |
| S | C or G |
| W | A or T |
| H | A, C, or T |
| B | C, G, or T |
| V | A, C, or G |
| D | A, G, or T |
| N | A, C, G, or T |
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Custom DNA & RNA Oligos service ships orders in as few as 4 business days depending on the modifications or purifications requested. Please refer to your order for specific turnaround time information.
Oligos Retained for Sanger will be available for sequencing within 1-2 business days depending on the time the order is placed.
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Custom DNA & RNA Oligo orders can only be delivered in dry format.
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If primers arrive dry, we recommend briefly centrifuging the tube to ensure that the DNA is at the bottom of the tube. Resuspend the sample in a volume or concentration appropriate for your needs and vortex briefly to mix well. Let the sample sit for 1-2 minutes. Before use, vortex briefly again.
*Please note the reconstitution solution is dependent on downstream applications and that TE may interfere with various enzymatic reactions.
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Dry oligos are stable and can be stored at -20°C for 1 year. Resuspended primers at higher concentrations (≥100 µM, in TE or water) are stable for 6 months at -20ºC.
*Please note that repeated freezing and thawing will reduce stability and shelf life.
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| Purification | Lengths (nt) | Purity | Recommended Uses |
|---|---|---|---|
| Desalted | 5-90 | >70% | Standard sequencing and PCR primers |
| PAGE | 11-150 | >90% | Long oligos |
| HPLC | 5-90 | >95% | Modified oligos or oligos used in sensitive applications |
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GENEWIZ synthesizes DNA oligos on a per-scale basis, which refers to the initial amount of material used in synthesis. Due to the inefficiency of the coupling reactions, yields are never equal to the initial amount of starting material. Yield is typically impacted by the nature and content of the sequence. Purification and modifications will also reduce the final yield delivered.| Length (nt) | Scale | Purification | Target Yield* |
|---|---|---|---|
| 5-90 | 25 nmol | Desalted | 5 nmol |
| HPLC | 2 nmol | ||
| 50 nmol | Desalted | 10 nmol | |
| HPLC | 4 nmol | ||
| PAGE | 2 nmol | ||
| 100 nmol | Desalted | 20 nmol | |
| HPLC | 8 nmol | ||
| PAGE | 4 nmol | ||
| 200 nmol | Desalted | 50 nmol | |
| HPLC | 20 nmol | ||
| PAGE | 10 nmol | ||
| 1 µmol | Desalted | 300 nmol | |
| HPLC | 120 nmol | ||
| PAGE | 60 nmol | ||
| 91-150 | 100 nmol | PAGE | 2 nmol |
| 200 nmol | PAGE | 5 nmol | |
| 1 µmol | PAGE | 25 nmol |
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| Dye | Max Ex (nm) | Max Em (nm) | Color | Compatible Quenchers |
|---|---|---|---|---|
| 6-FAM | 495 | 520 | Green | BHQ-1, BHQ-2, ECLIPSE, TAMRA, Dabcyl |
| JOE | 520 | 548 | Orange | BHQ-1, ECLIPSE, Dabcyl, TAMRA |
| TET™ | 521 | 536 | Orange | BHQ-1, BHQ-2, ECLIPSE, TAMRA, Dabcyl |
| HEX™ | 520 | 548 | Pink | BHQ-1, BHQ-2, ECLIPSE, TAMRA, Dabcyl |
| TAMRA | 544 | 576 | Rose | BHQ-2, ECLIPSE, Dabcyl |
| ROX™ | 576 | 601 | Red | BHQ-2, ECLIPSE |
| Cy®5 | 650 | 670 | Violet | BHQ-2 |
| Cy®3 | 550 | 570 | Red | BHQ-2, ECLIPSE |
| TexasRed® | 583 | 603 | Red | BHQ-2, ECLIPSE |
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GENEWIZ offers several options for synthesis and cost depends on the synthesis type, length, scales, modifications, and purifications required. Please contact dnaseq@azenta.com or +1-877-436-3949 ext. 2 with pricing inquiries or reach out directly to your Sales Representative.
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Yes, oligos containing RNA, LNA, 2-O-Me-RNA and other modified RNA nucleotides are available through our Custom Synthesis Service. Please submit an inquiry online through your GENEWIZ account.