牛白血病病毒PCR特异性引物的选择

U. Z. Kuzhebayeva, I. Beishova, V. Ulyanov, Т. V. Ulyanova, Alexandr Kovalchuk, N. Ginayatov, А. Z. Sidarova
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引用次数: 0

摘要

聚合酶链反应(PCR)方法被广泛用于解决各种问题。PCR广泛应用于细菌和病毒病原体的检测。引物是PCR非常重要的组成部分,因为扩增的特异性主要取决于引物。它们是酶发挥作用所必需的,并且对感兴趣的片段具有特异性。根据从NCBI网站上的国际数据库中选择的基因组核苷酸序列或病毒RNA单个片段的结果,鉴定出了大量的牛白血病病毒序列,这些序列存储在基因库中,并每日更新新数据。根据需要的参数,利用各种计算机程序进行引物的构建,主要有MUSCLE、UGENE V.36.0、Primer-BLAST、Oligo Analyzer等。然后根据设备附带的说明,在speedy 8909寡核苷酸合成器上合成设计的引物。通过实验,我们选择并合成了特异性的合成寡核苷酸env (g51)_1和env (g51)_2,用于建立牛白血病的PCR。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
SELECTION OF SPECIFIC PRIMERS FOR PCR IN BOVINE LEUKEMIA VIRUS
the polymerase chain reaction (PCR) method is widely used to solve various problems. PCR is widely used for the detection of bacterial and viral pathogens. Primers are a very important component of PCR, since the specificity of amplification depends primarily on them. They are necessary for the enzyme to work and are specific to the fragment of interest. Based on the results of the selection of nucleotide sequences of the genome or individual fragments of the virus RNA from the international database on the NCBI website, a large number of sequences for the bovine leukemia virus were identified, which are stored in gene banks and updated daily with new data. The construction of primers in compliance with the necessary parameters is carried out using various computer programs, the main of which are MUSCLE, UGENE V.36.0, Primer-BLAST, Oligo Analyzer and others. The designed primers were then synthesized on the Expedite 8909 oligonucleotide synthesizer, according to the instructions attached to the device. As a result of our experiments, we selected and synthesized specific synthetic oligonucleotides env (g51)_1 and env (g51)_2, for setting up PCR for bovine leukemia.
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