对于常见的缓冲分子而言,DNA吸收剂:在SELEXEX中可能存在缓冲干扰
Yuzhe Ding1, Ziyu Zhang1, Yunus A Kaiyum2
1Department of Chemistry, Waterloo Institute for Nanotechnology, University of Waterloo, Waterloo, Ontario, N2L 3G1, Canada. liujw@uwaterloo.ca.
Organic & biomolecular chemistry
|September 24, 2024
概括
研究人员开发出了能与Tris缓冲剂结合的aptamer,这是一个常见的实验室化学物质. 然而,试图为HEPES缓冲器创建aptamer的尝试没有成功,这突出了aptamer选择实验中的潜在挑战.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 适应物选择实验通常使用高度 (10-50mM) 的缓冲分子,而不是目标分子 (低μM).
- 这种度差异引发了人们对阿普坦体结合缓冲分子的可能性的担忧,特别是在失败的选择实验中.
研究的目的:
- 为了研究选择aptamers与常见的缓冲分子的可行性.
- 评估缓冲结合的阿帕特马可能干扰阿帕特马选择结果的可能性.
主要方法:
- 使用Tris和HEPES作为目标分子进行了aptamer选择.
- 选择的Tris胺体的结合亲和力是使用提奥夫拉T (ThT) 光光谱学和异热定位热量计 (ITC) 进行的.
- 核磁共振 (NMR) 谱学被用来确认阿普坦 - 标结合.
主要成果:
- 对Tris缓冲器特有的aptamers被成功分离出来.
- 特里斯亚胺体表现出2.9mM (ThT) 和43μM (ITC) 的解离常数 (Kd).
- 试图为HEPES缓冲器生成aptamers的尝试没有成功.
结论:
- 有可能选择能够结合像Tris.这样的缓冲分子的aptamer.
- 这项研究强调了在aptamer选择协议中考虑缓冲组合的重要性.
- 提供了关于使用特定缓冲器来减轻潜在的选择偏差的建议.
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