O6-Me-dG (iBu) CE-Phosphoramidite
O6-Me-dG (iBu) CE-Phosphoramidite
Key features
Show- Useful for mutagenic studies involving methylation.
- Classical isobutyryl protection.
Product information
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.
Applicable Products
LK2018 | O6-Me-dG-CE Phosphoramidite |
LK2019 | N6-Me-dA-CE Phosphoramidite |
LK2025 | O4-Me-dT-CE Phosphoramidite |
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.
Item |
Mol. Formula |
Mol. Wt. |
Unit Wt. |
250mg |
500mg |
1g |
LK2018 | C45H56N7O8P | 853.97 | 343.24 | 2.93 | 5.86 | 11.71 |
LK2019 | C41H50N7O6P | 767.87 | 327.24 | 3.26 | 6.51 | 13.02 |
LK2025 | 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
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.
Access support
Need some support with placing an order, setting up an account, or finding the right protocol?
Contact us