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Affinity Plus™ DNA & RNA Oligonucleotides

Oligos incorporating locked nucleic acids for enhanced stability and nuclease resistance

Affinity Plus DNA & RNA Oligonucleotides are custom, single-stranded, and duplexed sequences that contain 1−20 locked nucleic acid nucleotides. Incorporation of the locked nucleic acid nucleotides offers increased target specificity and oligo stability [1]. Refer to our Locked nucleic acids technology page to see the structure of these modified bases and to learn about the many applications they can facilitate.

Ordering

  • Cost-efficient, reliable oligos for a variety of hybridization-based applications
  • Helps you manage sequence melting temperature, resulting in greater hybridization specificity
  • Locked nucleic acids can improve stability and exonuclease resistance (in vitro and in vivo)

Affinity Plus Single-Stranded DNA

Shipped dry, or resuspended to your specifications.

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Affinity Plus Duplexed DNA

2 oligos, annealed and delivered in a single tube. Shipped dry.

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The following annealing fees will be applied to each duplex ordered:

100 nmol:

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250 nmol:

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1 µmol:

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Affinity Plus Single-Stranded RNA

Shipped dry.

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Affinity Plus Duplexed RNA

2 oligos, annealed and delivered in a single tube. Shipped dry.

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Annealing fees will be applied to duplexed oligos (per duplex), as follows:

100 nmol:

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250 nmol:

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1 µmol:

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Affinity Plus Single-Stranded DNA

Shipped dry, or resuspended to your specifications. A minimum of 24 and 96 oligos required for 96- and 384-well plates, respectively.

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Affinity Plus Duplexed DNA

1 oligo pair per well, annealed. Shipped dry. A minimum of 24 and 96 oligo duplexes are required for 96- and 384-well plates, respectively.

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The following annealing fee will be applied to each plate of duplexed DNA:

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For ordering inquiries, please contact us.

Affinity Plus Single-Stranded RNA

Shipped dry or resuspended to your specifications. A minimum of 24 or 96 oligos required for 96- or 384-well plates, respectively.

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For ordering inquiries, please contact us.

Product details

Affinity Plus DNA & RNA Oligonucleotides are locked nucleic acids containing single- and double-stranded sequences. With the Oligo Entry ordering tool, you can design the sequence you require with 1−20 locked nucleic acid nucleotides to suit your research needs.

Every Affinity Plus DNA & RNA Oligo you receive is deprotected and desalted to remove small molecule impurities. In addition, the identity of your oligos will be confirmed via proprietary ESI-mass spectrometry methods* and quantified by UV spectrophotometry to provide accurate yield measurements.

*With the exception of mixed base oligos, which could potentially represent multiple sequences therefore cannot accurately be evaluated by ESl mass spectrometry.

Using the Oligo Entry tool

To enter individual Affinity Plus nucleotides, insert a "+" before the base (+A, +C, +G, +T).

Product data

Affinity Plus oligonucleotides increase stability

Affinity Plus Oligonucleotides provide identical annealing properties as other manufacturers' locked nucleic acid sequences (Figure 1). Oligonucleotides that have locked nucleic acids have enhanced stability, increased melting temperatures, and higher nuclease resistance, relative to unmodified oligonucleotides.

Figure 1. Sequences containing Affinity Plus oligonucleotides show identical annealing properties to locked nucleic acid sequences from another vendor. The percentages of duplex melted base pairs are plotted as a function of temperature. The 15-mer sequence, GGTCCT+T+A+CTTGGTG, was synthesized with either Affinity Plus or other locked nucleic acid modifications incorporated at the +T, +A, and +C sites. These oligos were mixed with the complementary DNA strand (1 µM each strand) in 1 M Na+ buffer (pH 7). Melt curves were performed as described in Owczarzy, et al. [2]. The plots of duplex melted base pairs were averaged from at least 7 heating and cooling melting curves. Melting temperature was measured to be 67.7 ± 0.3 °C for the Affinity Plus oligo and 67.9 ± 0.3°C for the other locked nucleic acid oligo. Free energy of duplex hybridization at 37°C was determined to be -18.1± 0.9 kcal/mol for the Affinity Plus oligo and -18.7 ± 0.9 kcal/mol for the other locked nucleic acid oligo. These results illustrate the identical annealing properties of both sources of locked nucleic acid sequences.

Target identification in qPCR and SNP assays

The Affinity Plus sequences can be used as probes, for example, in qPCR and genotyping assays. Relative to unmodified oligonucleotides, the Affinity Plus RNA and DNA oligos have an increased affinity for targeted regions. Further the incorporation of Affinity Plus into these oligos makes them more stable than unmodified oligonucleotides [1]. This increased stability also makes it possible to use shorter probe designs, which can be helpful when target regions that are of a limited size [1].

For these applications, you can obtain probes as Affinity Plus qPCR probes. These Affinity Plus qPCR probes provide the same precise amplification as other locked nucleic acid probes and show more rapid target amplification over unmodified probes (see Figure 2 on the Product data tab of the Affinity. Plus qPCR Probes products page).

Frequently asked questions

References

  1. Kurreck J, Wyszko E, Gillen C, Erdmann VA. Design of antisense oligonucleotides stabilized by locked nucleic acids. Nucleic Acids Res. 2002;30(9):1911-8. doi:10.1093/nar/30.9.1911
  2. Owczarzy R, You Y, Moreira BG, et al. Effects of sodium ions on DNA duplex oligomers: improved predictions of melting temperatures. Biochemistry. 2004;43(12):3537-54. doi:10.1021/bi034621r
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