DNA Ligase Connects Two Fragments Having Exceptionally Short Complementary Termini at High Temperatures.

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ID: 84056
2020
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Abstract
DNA ligase ( DNA ligase) is widely employed for cloning, enzymatic synthesis, and molecular diagnostics at high temperatures (e.g., 65 °C). It has been long believed that the complementary ends must be very long (e.g., >30 bp) to place two DNA fragments nearby for the ligation. In the current study, the length of the complementary portion was systematically varied, and the ligation efficiency was evaluated using the high resolution melting (HRM) method. Unexpectedly, very short oligonucleotides (7-10 nt) were successfully ligated on the complementary overhang attached to a dsDNA at 70 °C. Furthermore, sticky ends with the overhang of only 4 nt long, available after scission with many restriction enzymes, were also efficiently ligated at 45-70 °C. The ligation yield for the 6-nt-long sticky ends was as high as 80%. It was concluded that DNA ligase can be used as a unique tool for DNA manipulation that cannot be otherwise easily accomplished.
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wang2020biochemistry Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Wang, Jing;Zhu, Jianming;Wang, Chenru;Zhou, Guangqing;Yu, Xin;Fan, Huijun;An, Ran;Komiyama, Makoto;Liang, Xingguo;
Journal Biochemistry
Year 2020
DOI
10.1021/acs.biochem.9b00866
URL
Keywords
diagnosis leishmania donovani visceral leishmaniasis high resolution melting hsp70 gene leptomonas seymouri melting peak

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