Novel Strategies to Construct Complex Synthetic Vectors to Produce DNA Molecular Weight Standards |
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Authors: | Zhe Chen Jianbing Wu Xiaojuan Li Chunjiang Ye He Wenxing |
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Institution: | (1) National Clinical Research Base of Traditional Chinese Medicine, Zhejiang Hospital of Traditional Chinese Medicine, Zhejiang Chinese Medical University, Hangzhou, 310006, People’s Republic of China;(2) Institute of Biotechnology, Zhejiang University, Hangzhou, 310029, People’s Republic of China;(3) Department of Biotechnology, College of Medicine and Life Sciences, University of Jinan, Jinan, 250022, People’s Republic of China;(4) State Key Laboratory of Microbial Technology, College of Life Science, Shandong University, Jinan, 250100, People’s Republic of China; |
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Abstract: | DNA molecular weight standards (DNA markers, nucleic acid ladders) are commonly used in molecular biology laboratories as
references to estimate the size of various DNA samples in electrophoresis process. One method of DNA marker production is
digestion of synthetic vectors harboring multiple DNA fragments of known sizes by restriction enzymes. In this article, we
described three novel strategies—sequential DNA fragment ligation, screening of ligation products by polymerase chain reaction
(PCR) with end primers, and “small fragment accumulation”—for constructing complex synthetic vectors and minimizing the mass
differences between DNA fragments produced from restrictive digestion of synthetic vectors. The strategy could be applied
to construct various complex synthetic vectors to produce any type of low-range DNA markers, usually available commercially.
In addition, the strategy is useful for single-step ligation of multiple DNA fragments for construction of complex synthetic
vectors and other applications in molecular biology field.
Zhe Chen and Jianbing Wu contributed to this work equally. |
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Keywords: | Complex synthetic vectors Molecular weight standards PCR screening of ligation products Sequential ligation End primer |
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