Processing
TruGrade™ DNA Oligos
Reduce barcode crosstalk in your multiplexed NGS experiments
Quality requirements for synthetic oligos used in next generation sequencing (NGS), such as indexed adapters and barcoded fusion primers, have reached unprecedented heights. While traditional purification methods (e.g., HPLC) can effectively increase the amount of full-length oligo, they are incapable of reducing cross-contamination that can occur in co-synthesized oligos. In contrast to applications where such oligo-to-oligo crosstalk has less impact, the use of indexed adapters containing even low levels of cross-contamination can lead to barcode misalignment and sample misidentification, which may confound experiments attempting to identify rare variants.
Ordering
TruGrade oligos are manufactured by proprietary methods that have been shown to reduce index crosstalk and increase success of multiplex NGS experiments. With TruGrade oligos, you can:
- Obtain DNA oligos with cross-contamination levels as low as 0.01%
- Select from a variety of product options to suit your requirements
- Experience the quality and turnaround time for which IDT is known
DNA oligos
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Ultramer™ DNA oligos
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DNA oligos
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Ultramer DNA oligos
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Product details
TruGrade DNA Oligos
With rapidly increasing throughput of sequencing platforms, sample multiplexing has become a common solution to economically address new sequencing capacities. However, pooling indexed samples into one sequencing run comes with the risk of sample misidentification. While this can be attributed to many experimental factors, crosstalk during oligo synthesis is a major contributor. TruGrade DNA oligos are manufactured using an exclusive production process that reduces the risk of oligonucleotide crosstalk and thereby minimizing the risk of barcode misalignment during multiplexing NGS applications.
The TruGrade synthesis and processing service is available for HPLC-purified DNA oligos and Ultramer™ DNA oligos. They are available in plates and tubes and can be formulated to your specifications. TruGrade DNA oligos can be used in a variety of applications including, but not limited to:
- Library preparation that incorporates barcoded adapters for multiplexing
- PCR using barcoded fusion primers for multiplex amplicon sequencing
- Other molecular biology applications that are sensitive to oligo crosstalk
Specifications
| Parameter | HPLC purified DNA oligos | Ultramer DNA Oligos |
|---|---|---|
| Typical cross-contamination1 | 0.10–0.50% | 0.01–0.05% |
| Synthesis scale | 100 nmol, 250 nmol, or 1 µmol | N/A |
| Yield delivered | Variable | 4 nmol or 20 nmol |
1 Typical cross-contamination levels were determined using in-house proprietary QC techniques and information from our customers.
Modifications
| Modification type | Modification | DNA oligos (100 nmol, 250 nmol, 1 µmol) | Ultramer DNA Oligos (4 nmol, 20 nmol) |
|---|---|---|---|
| 5′ modifications | 5′ 5-Methyl dC | • | • |
| 5′ Amino Modifier C12 | • | ||
| 5′ Amino Modifier C6 | • | ||
| 5′ Biotin | • | • | |
| 5′ deoxyInosine | • | • | |
| 5′ deoxyUridine | • | • | |
| 5′ Phosphorylation | • | • | |
| Internal modifications | Int 5-Methyl dC | • | • |
| Int deoxyInosine | • | • | |
| Int deoxyUridine | • | • | |
| Int Spacer 18 | • | • | |
| 3′ modifications | 3′ Amino Modifier | • | • |
| 3′ ddC | • | ||
| 3′ C3 Spacer | • | ||
| 3′ Phosphorylation | • | ||
| Modified bases and bonds | 2′ O-Methyl RNA bases | • | • |
| Phosphorothioate Bond | • | • |
Resources
RUO22-1141_001