Cannot add to favourites
An option must be selected in order to add to favourites
Methylating agents are common carcinogens which function by methylation of nucleobases in DNA. Cellular polynucleotides are alkylated by endogenous components, such as S-adenosylmethionine. Chemically, SN1 agents such as alkylnitrosourea and N-alkyl-N-nitro-N-nitrosoguanidine will react with the N7 position of guanine, N3 of adenine, O6 of guanine, O2 or O4 of pyrimidines, and the non-phosphodiester oxygen atoms of the phosphate backbone. In contrast, SN2 chemical agents such as methyl methanesulfonate and dimethyl sulfate react primarily with the N1 position of adenine (1-Methyl-2'-deoxyadenosine) and N3 of cytosine. To examine some of these resulting mutagenic effects, the methylated products O6-Me-dG- CE Phosphoramidite, N6-Me-dA-CE Phosphoramidite, and O4-Me-dT-CE Phosphoramidite can be incorporated in oligonucleotides. To avoid chain branching during synthesis when using DCI as activator, N6-Me-dA is offered with acetyl protection. O6-Me-dG- CE Phosphoramidite has classic iBu protection.
Physical & Dilution Data
Dilution volumes (in ml) are for 0.1M solutions in dry acetonitrile (LK4050). Adjust accordingly for other concentrations. For µmol pack sizes, products should be diluted as 100µmol/ml to achieve 0.1M, regardless of molecular weight.
No changes are required from the standard method recommended by the synthesiser manufacturer. Coupling is as per standard nucleoside amidites.
Cleavage & Deprotection
LK2018 and LK2025 -
- Empty the synthesis resin from the column into a sample tube.
- 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.
- Place the tube in a dry dark place and leave it for 5 days.
- Reduce to small volume.
- Add 1ml of 10mM aqueous sodium hydroxide solution and desalt or purify the oligonucleotide using standard procedures.
LK2019 - 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.