The factors affecting the reproducibility of micro-volume DNA mass quantification in Nanodrop 2000
The factors affecting the reproducibility of micro-volume DNA mass quantification in Nanodrop 2000
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Nanodrop 2000分光光度计微量DNA质量定量重现性的影响因素
DOI：
10.1016/j.ijleo.2017.08.031
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发表时间：
2017-01-01
期刊：
影响因子：
3.1
通讯作者：
Li, Wenjie
中科院分区：
文献类型：
--
作者：
Yu, Songcheng;Wang, Yan;Li, Wenjie
在进行各种DNA操作和分子分析之前，应准确估算DNA质量浓度。然而，关于影响微量DNA质量定量重现性的因素尚无系统报道。在本研究中，对微量分析仪的性能以及影响DNA定量的因素进行了系统研究。结果表明，Nanodrop 2000分光光度计在30 - 500 ng/μL的浓度范围内具有良好的精密度和准确度。DNA溶液的均一性对定量结果有很大影响。储存后DNA溶液不同部位的浓度顺序为底部＞中部＞上部。此外，随着延迟时间的延长，测量结果会增加。较小上样量的增加速度比上样量较大时更快。另外，高浓度上样样品的残留对下一次低浓度样品的测定影响较小。总之，Nanodrop 2000分光光度计对于微量样品的DNA定量是可靠的。在测定过程中应注意溶液的均一性、延迟时间、上样样品体积以及交叉污染。这可作为微量DNA定量的参考指南。（C）2017爱思唯尔有限公司。保留所有权利。
The DNA mass concentration should be accurately estimated prior to various DNA manipulations and molecular analyses. However, there was no systematic report about the factors affecting the reproducibility of micro-volume DNA mass quantification. In this study, the performance of micro-volume analyzer and the factors affecting DNA quantification were studied systematically. The results revealed that the precision and accuracy of Nanodrop 2000 spectrophotometer were excellent within concentration range from 30 to 500 ngipl. The homogeneity of DNA solution had great effect on the quantification result. The concentration order for different parts of DNA solution after storage was bottom > center > upper. Furthermore, the measured results increased as delay time extended. The increased speed for smaller loading volume was faster than that for larger loading volume. In addition, the residue from the loaded sample of high concentration had a little effect on the next determination for low concentration sample. In summary, Nanodrop 2000 spectrophotometer was reliable for DNA quantification of micro-volume sample. And attention should be paid to solution homogeneity, delay time, loading sample volume and cross contamination during determination. This could be referred as guidance for micro-volume DNA quantification. (C) 2017 Elsevier GmbH. All rights reserved.