| | Repair and mutagenic potency of 8-oxoG:A and 8-oxoG:C base pairs in mammalian cells -- Le Page et al. 26 (5): 1276 -- ... |
 | | Samples (20 µg) of the gapped duplex plasmid DNA formed, were hybridized overnight with 5 µg of the 5[prime] |
 | | P-phosphorylated GO carrying oligonucleotide (GATC GGC GCC GGOC GGT GTG) in T4 ligation buffer containing 1 mM ATP, then ligated for 20 min with 400 U T4 DNA ligase (Biolabs) at 12°C. Finally, covalently closed molecules were purified by isopycnic centrifugation on an ethidium bromide/cesium chloride gradient. |
 | | Fractions collected from the gradient were analyzed on a 1% agarose gel and those corresponding to the closed circular double-stranded DNA vector were pooled, dialyzed against TE (1 mM Tris pH 7.8, 1 mM EDTA) and then ethanol precipitated. |
| nar.oxfordjournals.org /cgi/content/full/26/5/1276 (5321 words) |