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Optimisation and evaluation of Human Leucocyte Antigen-G1 expression in Placental Tissue using Real Time Polymerase Chain Reaction

机译:实时聚合酶链反应优化和评估胎盘组织中人白细胞抗原-G1的表达

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Laboratory diagnostic techniques serve as essential tools for screening diseases, and monitoring clinical outcomes. Using this as motivation, this study optimised the Real time Quantification Polymerase Chain Reaction (RT-qPCR) technique to specifically quantify Human Leucocyte Antigen-G1 expression in placental tissue. Synthesis of complementary deoxyribose nucleic acid (cDNA) was performed using ribose nucleic acid (RNA) extracted from human placental tissue. Amplification of cDNA, using specifically designed primers, complementary to the full length HLA-G1 isoform, was performed using RT-qPCR. Data analysis was concluded using the “second derivative method’’ (Roche Molecular Biochemicals Light Cycler Relative Quantification Software, Version 3.5). Amplification of cDNA was successfully accomplished. Samples with HLA-G1 transcripts gave a fluorescent signal for the 98-base pair (bp) PCR product, characterized by a melting peak at 86.5?C. The RT-qPCR assay is an efficient and reliable tool, readily adaptable for quantifying HLA- G1 expression in placental tissue, to ensure normal full term pregnancies.
机译:实验室诊断技术是筛查疾病和监测临床结果的重要工具。以此为动机,本研究优化了实时定量聚合酶链反应(RT-qPCR)技术,以特异性定量胎盘组织中人白细胞抗原G1的表达。使用从人胎盘组织提取的核糖核酸(RNA)进行互补的脱氧核糖核酸(cDNA)的合成。使用专门设计的引物,与全长HLA-G1同工型互补,使用RT-qPCR扩增cDNA。数据分析是通过“二阶导数法”(Roche Molecular Biochemicals Light Cycler相对定量软件,版本3.5)得出的。 cDNA的扩增成功完成。具有HLA-G1转录本的样品对98个碱基对(bp)的PCR产物发出荧光信号,其特征是在86.5°C时有一个熔解峰。 RT-qPCR分析是一种有效且可靠的工具,可轻松用于定量胎盘组织中HLA-G1的表达,以确保正常的足月妊娠。

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