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Real Time RT-PCR02:57

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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.
The real-time quantification of the number of amplified products is...
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快速和灵敏的细菌活性的量化,使用电比计光传感.

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使用GelRed和SYTO 9的新型比度光方法可以有效地区分活菌和死菌. 这种敏感的技术通过准确评估细菌生存能力,有助于食品安全和药物开发.

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科学领域:

  • 微生物学 微生物学
  • 生物技术是生物技术.
  • 分析化学 分析化学

背景情况:

  • 细菌活力评估对于食品安全和抗菌研究至关重要.
  • 传统的可行性测试方法在灵敏度和准确度上可能受到限制.

研究的目的:

  • 开发一种新,灵敏和准确的比度光法,用于细菌生存能力的评估.
  • 建立一种使用DNA结合染料区分活菌和死菌的方法.

主要方法:

  • 使用GelRed作为DNA结合染料,并指出活细菌排除它,而死细菌吸收它.
  • 采用子来最大限度地减少GelRed的非特异性吸附.
  • 使用SYTO 9 (一种膜透性染料) 开发了一种双染色技术.
  • 建立了一种定量计光试验,用于量化死亡细菌.

主要成果:

  • 凝红/SYTO 9方法表现出高灵敏度,检测到死亡细菌在埃舍里奇亚大肠杆菌O157:H7.7.中的水平低至0.1%.
  • 观察到一个强的线性相关性 (R2 = 0.97) 的比度光和理论死亡的细菌比率.
  • 该方法在评估热处理的杀菌效果方面被证明是有效的,在50°C时显示细菌死亡.

结论:

  • 与传统的可行性测定相比,开发的比度光方法提供了更高的灵敏度,准确性和安全性.
  • 这种技术在微生物研究,病原体检测和药物开发方面具有广泛的应用.
  • 该方法显示了适应哺乳动物细胞活力研究的潜力.