双向电泳法分离样品中单、双链DNA
Separating single-stranded DNA and double-stranded DNA in samples by bidirectional electrophoresis
摘要
目的：探索双向电泳法分离复杂样品样品中单、双链DNA片段的方法。方法：采用双向电泳法，通过垂直电泳和水平电泳，并调整水平电泳的温度、功率、电泳时间实现分离复杂样品中的单、双链DNA。结果：在常温下进行水平电泳发现没有出现单双链DNA片段分离的现象，推测出现凝胶内复性，导致两个方向均存在双链DNA片段。在55℃下进行水平电泳，当条件为5W恒功率，时间45分钟时，不能达到分离所需的迁移速率，需进一步增加电泳功率并延长电泳时间。结论：垂直电泳后，凝胶92℃加热3分钟，55℃下恒功率15W水平电泳80分钟可实现单双链DNA片段的分离。
Abstract
Purpose: To investigate the methods for isolating single- or double- stranded DNA fractions in complex samples. Approach: The single- and double-stranded DNA in complex samples was separated by bidirectional electrophoresis, and the separation was realized by adjusting the temperature, power and electrophoresis time of horizontal electrophoresis. Results：Horizontal electrophoresis at room temperature showed that there was no separation of single and double-stranded DNA fragments, and it was speculated that in-gel renaturation of ssDNA fragments led to the presence of dsDNA fragments in both directions. Horizontal electrophoresis is performed at 55 C, when the condition is 5W constant power for 45 minutes, the migration rate required for separation cannot be achieved, and it is necessary to further increase the electrophoresis power and extend the electrophoresis time.Consequence: After vertical electrophoresis, the gel was heated at 92 C for 3 minutes, and the separation of single and double-stranded DNA fragments could be realized by horizontal electrophoresis of 15W at 55 C for 80 minutes.关键词
双向电泳/单链DNA/双链DNA/分离Key words
bidirectional electrophoresis/single-stranded DNA/double-stranded DNA/sepatate引用本文复制引用
邱新宇,崔凤梅,刘一桐,陈秋.双向电泳法分离样品中单、双链DNA[EB/OL].(2023-03-13)[2026-04-05].http://www.paper.edu.cn/releasepaper/content/202303-137.学科分类
生物科学研究方法、生物科学研究技术/分子生物学
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