| ATG Trimer Phosphoramidite |
Catalog Number: 10-1032-xx
Description: O4-Me-dT-CE Phosphoramidite
5'-Dimethoxytrityl-O4-methyl-2'-deoxyThymidine, 3'-[(2-cyanoethyl)-
(N,N-diisopropyl)]-phosphoramidite |
| Formula: C41H51N4O8P |
M.W.: 758.85 |
F.W.: 318.22 |
Diluent: Anhydrous Acetonitrile |
| Coupling: No changes needed from standard method recommended by synthesizer manufacturer. Monomers that allow for UltraMILD deprotection are recommended (Catalog Numbers: dA: 10-1601-xx, dC: 10-1015-xx, dG: 10-1621-xx, dT: 10-1030-xx.) To avoid any exchange of the iPr-Pac group on the dG with acetyl, use the UltraMild Cap Mix A (40-4210-xx/ 40-4212-xx). |
| Deprotection: If UltraMild Monomers were used: 0.05M Potassium Carbonate in Methanol, 4hrs at Room Temp.DMT-Off Synthesis: Add 6uL acetic acid per mL 0.05M potassium carbonate in methanol. Dry under vacuum. Desalt per standard procedures. DMT-On Synthesis: Use a Glen-Pak Purification Cartridge (60-5100-XX/60-5200-XX) to purify and desalt the oligonucleotide. Dilute the 0.05M potassium carbonate in methanol with 100mg/mL Sodium Chloride to a final concentration of 1:4(v/v). Load onto the prepped Glen-Pak Cartridge and purify as normal. If Standard Monomers were used: 1) Prepare a solution of 10% DBU in anhydrous methanol. DBU (1,8-Diazabicyclo[5.4.0]undec-7-ene) is available from Aldrich (139009-25G) as is anhydrous methanol (322415-100ML). 2) Transfer the CPG to a clean, dry glass vial and add 1mL of the 10% DBU solution. 3) Seal the vial and leave for 5 days in the dark at room temperature. 4) Transfer the contents to a micro centrifuge tube and reduce under vacuum to a small volume. 5) Add 1mL of 10mM aqueous sodium hydroxide solution and desalt or purify the oligonucleotide using standard procedures. Technical Bulletin |
| Storage: Refrigerated storage, maximum of 2-8°C, dry |
| Stability in Solution: 2-3 days |
MUTAGENESIS
Cellular polynucleotides are alkylated by endogenous components, such as S-adenosylmethionine, or after reacting with two general classes of environmental and laboratory chemicals. SN1 chemical agents include alkylnitrosourea and N-alkyl-N-nitro-N-nitrosoguanidine that 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 avoid chain branching during synthesis when using DCI as activator, N6-Me-dA is offered with acetyl protection.
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