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相关概念视频

Real Time RT-PCR02:57

Real Time RT-PCR

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.
The real-time quantification of the number of amplified products is...

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A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies
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一个基于光的协议,用于定量定位CUT&RUN缓冲元件的缓冲元件.

Andrew Katznelson1, Kenneth Zaret1

  • 1Institute for Regenerative Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.

STAR protocols
|February 8, 2024
PubMed
概括

这项研究引入了一种基于光的方法,以优化在目标下切割和释放,使用核酶 (CUT&RUN) 缓冲条件. 该协议确保了有效的细胞透和表皮质保留,以便进行准确的基因组分析.

科学领域:

  • 分子生物学分子生物学
  • 基因组学就是基因组学.
  • 生物化学 生物化学

背景情况:

  • 使用核酶在目标下切割和释放 (CUT&RUN) 是绘制蛋白质-DNA相互作用和染色质中的基因组修饰的关键技术.
  • 优化缓冲条件对于高效的细胞通透性和在CUT&RUN试验中保持表皮质完整性至关重要.

研究的目的:

  • 开发和介绍一种基于光的协议,用于对CUT&RUN缓冲元件进行定量定位.
  • 为了优化细胞透和对象表皮质保留在染色质上,以提高CUT&RUN试验的性能.

主要方法:

  • 细胞被捕获在康卡纳瓦林A珠上.
  • 使用基于光的测定方法来定位CUT&RUN缓冲器中的数字和NaCl度.
  • 使用光成像来确定最佳的缓冲条件.

主要成果:

  • 开发的协议允许对CUT&RUN缓冲器组件进行定量评估.
  • 确定了数字和NaCl的最佳度,以增强细胞通透性和表皮质保留.
  • 基于光的定位为测试优化提供了一个强大的方法.

结论:

  • 这种基于光的定位协议提供了一种高效和定量方法来优化CUT&RUN试验.
关键词:
细胞生物学 细胞生物学基于细胞的测试在 ChIP-seqq.染色体免疫沉 (ChIP) 是一种遗传学 遗传学 是一个分子生物学分子生物学

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  • 优化的条件提高了鉴定蛋白质和基因组修饰的基因组结合位的可靠性和准确性.