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相关概念视频

RNA-seq03:21

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RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
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Sanger Sequencing

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DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
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Real Time RT-PCR02:57

Real Time RT-PCR

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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
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简单而实用的DNA量化方法用于DNA编码图书馆合成.

Hangke Ma1, Zhaomei Sun1, Lijun Xue1

  • 1Pharmaron (Ningbo) Technology Development Co., Ltd., No. 800 Bin-Hai Fourth Road, Hangzhou Bay New Zone, Ningbo 315336, China.

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概括

DNA编码库 (DEL) 合成涉及可能损害DNA的化学反应. 这项研究引入了一种新的LC-MS方法来准确量化DNA损伤,揭示了与qPCR等传统方法的显著差异.

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科学领域:

  • 药用化学 医学化学
  • 生物技术是生物技术.
  • 分析化学 分析化学

背景情况:

  • 用DNA编码的库 (DELs) 对于药物发现中的匹配发现至关重要.
  • 德尔合成的进步扩大了化学空间,但也引发了对DNA完整性的担忧.
  • 现有的DNA损伤分析 (qPCR,测序) 在准确性和一致性方面存在局限性.

研究的目的:

  • 开发和验证一种精确的方法来量化DEL合成期间的DNA损伤.
  • 评估常见的DNA化学反应对DNA完整性的影响.
  • 将新方法的结果与传统的DNA损伤评估技术进行比较.

主要方法:

  • 使用液体染色学-质谱法 (LC-MS) 开发一种外部标准方法.
  • 应用LC-MS方法来评估来自各种DNA化学反应的DNA损伤.
  • 对LC-MS数据与传统定量PCR (qPCR) 测量进行比较分析.

主要成果:

  • 该LC-MS方法提供了在DEL合成过程中DNA降解的准确和实际量化.
  • 在LC-MS方法和传统的qPCR评估之间观察到显著的分歧.
  • 在DEL合成中常用的某些化学转化会导致大量的DNA损伤.

结论:

  • 开发的LC-MS方法提供了一种可靠的方法来监测DEL工作流中的DNA完整性.
  • 评估DELs中DNA损伤的传统方法可能会产生不一致或误导的结果.
  • 精确的DNA损伤评估对于优化DEL合成和确保数据可靠性至关重要.