Rotational dynamics of hexaamminecobalt(III) bound to oligomeric DNA: correlation with cation-induced structural transitions.
Rotational dynamics of hexaamminecobalt(III) bound to oligomeric DNA: correlation with cation-induced structural transitions.
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六氨合钴 (III) 与寡聚 DNA 结合的旋转动力学：与阳离子诱导的结构转变的相关性。
DOI：
10.1021/bi00095a002
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发表时间：
1993
期刊：
影响因子：
2.9
通讯作者：
Braunlin,WH
中科院分区：
文献类型：
--
作者：
Xu,Q;Jampani,SR;Braunlin,WH
材料和方法实验程序。所有寡核苷酸均在 Applied Biosystems PCR-Mate DNA 合成仪上以 10 mM 规模合成。合成后，将寡核苷酸溶解在缓冲液中，过滤除去有机盐，并使用 Spectrapore 1000 MW 截止透析管进行透析。至少 4 小时后进行透析改变。第一个变化是针对 0.1 M Tris，pH 7.6，然后是两次 0.1 M 氯化钠变化，然后是三次 0.001 M 氯化钠变化。寡核苷酸浓度是通过在变性条件下进行的紫外吸光度测量，使用最近邻法计算的消光系数来确定的（Cantor 等人，1970）。 DNA 样品的初始 Na+ 浓度通过将积分 23Na 信号强度与 D2O 中 NaCl 标准曲线的强度进行比较来确定。如前所述，合成寡核糖核苷酸 r (GGCCGGCC)，通过反相 HPLC 纯化，并透析到低盐溶液中 (Xu 等人，1993)。
MATERIALS AND METHODSExperimental Procedures. All oligonucleotides were synthesized on an Applied Biosystems PCR-Mate DNA syn-thesizer, on a 10-mM scale. Following synthesis, the oligo-nucleotides were dissolved in buffer, filtered to remove organic salts, and dialyzed using Spectrapore 1000 MW cutoff dialysis tubing. Dialysis changes were performed after at least 4 h. The first change was against 0.1 M Tris, pH 7.6, followed by two changes of 0.1 M NaCl and then three changes of 0.001 M NaCl. The oligonucleotide concentrations were determined from UV absorbance measurements, performed under de-naturing conditions, using extinction coefficients calculated by the nearest-neighbor method (Cantor et al., 1970). The initial Na+ concentrations of the DNA samples were determined by comparison of the integrated 23Na signal intensities with intensities from a standard curve of NaCl in D2O. The oligoribonucleotide r (GGCCGGCC) was synthesized, purified by reverse-phase HPLC, and dialyzed into low-salt solution as previously described (Xu et al., 1993).