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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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The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
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Protons and neutrons, collectively called nucleons, are packed together tightly in a nucleus. With a radius of about 10−15 meters, a nucleus is quite small compared to the radius of the entire atom, which is about 10−10 meters. Nuclei are extremely dense compared to bulk matter, averaging 1.8 × 1014 grams per cubic centimeter. If the earth’s density were equal to the average nuclear density, the earth’s radius would be only about 200 meters.
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在Agrostemma githago中的基因基因稳定性使用定量实时PCR.

Monika Bielecka1, Bartosz Pencakowski1, Marta Stafiniak1

  • 1Department of Pharmaceutical Biology and Biotechnology, Faculty of Pharmacy, Wroclaw Medical University, Borowska 211A, 50-556 Wroclaw, Poland.

International journal of molecular sciences
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概括

这项研究确定了Agrostemma githago中定量实时PCR (qPCR) 最稳定的参考基因. 推使用素H3 (H3) 和真核转化启动因子5A1 (TIF5A1-2) 进行精确的基因表达正常化.

关键词:
农业农业公司 githagogo在RefFinder中找到.家庭管理基因的基因qRT-PCR的正常化方法基因基因验证参考基因验证

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科学领域:

  • 分子生物学分子生物学
  • 植物科学 植物科学
  • 生物技术是生物技术.

背景情况:

  • 定量实时PCR (qPCR) 对于基因表达分析至关重要,需要验证的参考基因才能准确正常化.
  • 通常使用的参考基因在不同的植物组织,发育阶段和实验条件中显示出可变的表达.
  • 目前没有可用于Agrostemma githago (玉米) 的验证参考基因.

研究的目的:

  • 确定和验证最稳定的基因参考基因在Agrostemma githago基因表达研究.
  • 促进对自然产品生物合成和这种物种的特殊代谢调节的研究.

主要方法:

  • 根据文献和转录组数据,选出了七个候选家政基因.
  • 使用定量实时PCR (qRT-PCR) 来测量40个不同的Agrostemma githago样本中的转录水平.
  • 使用RefFinder平台评估表达稳定性,将geNorm,NormFinder,BestKeeper和Δ-Ct方法整合在一起.

主要成果:

  • 参考基因稳定性因特定器官,发育阶段和实验条件而异.
  • 在各种体外条件下,TIF5A1-2和GAPDH表达最稳定.
  • EF1α和H3在土壤中生长的植物的不同器官中优越.
  • 基因组H3 (H3) 和TIF5A1-2被确定为Agrostemma githago在所有测试条件中最稳定,最普遍的两个参考基因.

结论:

  • 该研究成功地确定并验证了Agrostemma githago最稳定的参考基因.
  • 建议在Agrostemma githago的qPCR研究中使用H3和TIF5A1-2进行准确的正常化.
  • 这些发现为未来的Agrostemma githago和相关物种的转录和功能研究提供了基础.