One of the major sources of DNA damage in all organisms is the UV component of sunlight. The predominant reaction induced by UV light on DNA is dimerization of adjacent pyrimidine bases leading to cyclobutane dimers (CPDs). The dimers formed in the most significant quantity are the cis-syn cyclobutane dimer of two thymine bases. Although formed routinely, these dimer products are efficiently excised and repaired enzymatically by nucleotide excision repair (NER) or the dimerization is reversed by photolase enzymes. A further mode of oxidative damage is radiation-induced damage of DNA, which has been shown to lead to bridged cyclonucleosides. The purines, cyclo-dA and cyclo-dG, are predominantly formed, although the cyclo pyrimidines have also been detected. Cyclo-dA is doubly intriguing since it contains both damaged base and damaged sugar residues and, as such, should have a considerable biological impact. In a manner analogous to thymine dimer, cyclo purines cause significant distortion of the regular DNA helix and these lesions are repaired not by base excision repair (BER) but by NER.
Details
Usage
Coupling: 10 minute coupling recommended.
Deprotection: |1. Remove the methyl phosphate groups by treating the support with thiophenol/triethylamine/THF (1:2:2) v/v/v for 45 minutes at room temperature. Wash the support with THF 1X, methanol 5X, acetonitrile 3X and dry with air or argon.|2. Cleave and deprotect the oligo in the dark with ammonium hydroxide at room temperature under conditions appropriate to remove the protecting groups on the nucleobases.
Specifications
Diluent
Anhydrous Acetonitrile
Storage
Freezer storage, -10 to -30°C, dry
Stability
2-3 days
Dilution/Coupling Data
The table below show pack size data and, for solutions, dilution and approximate coupling based on normal priming procedures.