Mutagenesis - Use of O6-Me-dG, N6-Me-dA and O4-Me-dT

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Applicable Products

2018 O6-Me-dG-CE Phosphoramidite
2019 N6-Me-dA-CE Phosphoramidite
2025 O4-Me-dT-CE Phosphoramidite

Physical & Dilution Data

Dilution volumes (in ml) are for 0.1M solutions in dry acetonitrile (4050). Adjust accordingly for other concentrations. For µmol pack sizes, products should be diluted as 100µmol/ml to achieve 0.1M, regardless of molecular weight.

Item

Mol. Formula

Mol. Wt.

Unit Wt.

250mg

500mg

1g

2018 C45H56N7O8P 853.97 343.24 2.93 5.86 11.71
2019 C41H50N7O6P 767.87 327.24 3.26 6.51 13.02
2025 C41H51N4O8P 758.85 318.22 3.29 6.59 13.18

Coupling

No changes are required from the standard method recommended by the synthesiser manufacturer. Coupling is as per standard nucleoside amidites.

Cleavage & Deprotection

2018 and 2025 -

  1. Empty the synthesis resin from the column into a sample tube.
  2. Add 900μl anhydrous MeOH and 100μl DBU (1,8-Diazabicyclo [5.4.0]undec-7-ene) to the resin in the sample tube and cap tightly.
  3. Place the tube in a dry dark place and leave it for 5 days.
  4. Reduce to small volume.
  5. Add 1ml of 10mM aqueous sodium hydroxide solution and desalt or purify the oligonucleotide using standard procedures.

2019 - Use AMA at room temperature for 2h. Employ dmf-G and Ac-C within the sequence.

Storage & Stability

All phosphoramidites are stored refrigerated at a maximum of 2-8°C. Stability in solution is 2-3 days.

 

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