The Efficiency of Various DNA Polymerases for Amplification of Long Sequences from Genomic DNA and cDNA of Cultivated Potatoes

IF 1 4区生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Applied Biochemistry and Microbiology Pub Date : 2023-08-12 DOI:10.1134/S0003683823040026

A. D. Antipov, N. E. Zlobin

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引用次数: 0

Abstract

Amplification of long fragments from complex templates, such as eukaryotic genomic DNA, is considered a difficult task for most DNA polymerases. In this research, six DNA polymerases were used to amplify full-length sequences from the genomic DNA of Solanum tuberosum genes encoding translation initiation factors of the eIF4E family, as well as for the synthesis of fragments of the potato Y virus genome from cDNA of potato plants infected by this virus. It was found that the efficiency of amplification by various DNA polymerases generally decreased with the increasing length of the amplicons. LongAmp and Platinum SuperFi II polymerases demonstrated the highest efficiency in the synthesis of long fragments, which made it possible to synthesize PCR products with a length of more than 10 000 base pairs with high efficiency. The lowest efficiency was demonstrated by Encyclo polymerase. None of the DNA polymerases provided efficient amplification of all the studied DNA fragments. At the same time, any of the studied DNA fragments could be effectively amplified using at least one DNA polymerase variant. Thus, the choice of DNA polymerase was of key importance for the efficiency of the synthesis of a desired PCR product.

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不同DNA聚合酶对栽培马铃薯基因组DNA和cDNA长序列扩增的效率

从复杂的模板中扩增长片段，如真核生物基因组DNA，对大多数DNA聚合酶来说是一项艰巨的任务。本研究利用6种DNA聚合酶从马铃薯马铃薯中扩增编码eIF4E家族翻译起始因子基因的基因组DNA全长序列，并从感染该病毒的马铃薯植株的cDNA中合成马铃薯Y病毒基因组片段。结果表明，随着扩增子长度的增加，各种DNA聚合酶的扩增效率普遍降低。LongAmp和Platinum SuperFi II聚合酶对长片段的合成效率最高，可以高效合成长度超过10000个碱基对的PCR产物。Encyclo聚合酶的效率最低。没有一种DNA聚合酶能有效地扩增所有研究的DNA片段。同时，任何研究的DNA片段都可以使用至少一种DNA聚合酶变体进行有效扩增。因此，DNA聚合酶的选择对于合成所需PCR产物的效率至关重要。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Applied Biochemistry and Microbiology 生物-生物工程与应用微生物

CiteScore

1.70

自引率

12.50%

发文量

审稿时长

6-12 weeks

期刊介绍： Applied Biochemistry and Microbiology is an international peer reviewed journal that publishes original articles on biochemistry and microbiology that have or may have practical applications. The studies include: enzymes and mechanisms of enzymatic reactions, biosynthesis of low and high molecular physiologically active compounds; the studies of their structure and properties; biogenesis and pathways of their regulation; metabolism of producers of biologically active compounds, biocatalysis in organic synthesis, applied genetics of microorganisms, applied enzymology; protein and metabolic engineering, biochemical bases of phytoimmunity, applied aspects of biochemical and immunochemical analysis; biodegradation of xenobiotics; biosensors; biomedical research (without clinical studies). Along with experimental works, the journal publishes descriptions of novel research techniques and reviews on selected topics.