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Efficient DNA Click Reaction Replaces Enzymatic Ligation.
Bioconjugate Chemistry ( IF 4.0 ) Pub Date : 2020-01-08 , DOI: 10.1021/acs.bioconjchem.9b00805 Michael Kollaschinski 1 , Jessica Sobotta 1 , Alexander Schalk 1 , Thomas Frischmuth 1 , Birgit Graf 1 , Sascha Serdjukow 1
Bioconjugate Chemistry ( IF 4.0 ) Pub Date : 2020-01-08 , DOI: 10.1021/acs.bioconjchem.9b00805 Michael Kollaschinski 1 , Jessica Sobotta 1 , Alexander Schalk 1 , Thomas Frischmuth 1 , Birgit Graf 1 , Sascha Serdjukow 1
Affiliation
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We report a chemical DNA-DNA ligation method based on copper-catalyzed azide-alkyne cycloaddition (CuAAC). We demonstrate that ion addition dramatically influences the efficiency of the so-called click reaction. Even without any further additions, such as typically splint oligonucleotides for preorganization, the "click ligation" yields up to ∼83% product without any byproducts. Additionally, purification of the desired product is straightforward. In comparison to enzymatic ligation methods used to introduce adapters into, e.g., mRNA library preparation, this targeted chemical ligation method exhibits several advantages: increased ligated product and no adapter or cDNA oligomers byproducts. The advantages of the click ligation method were demonstrated by incorporation of azide modified nucleotides by several enzymes as well as broad polymerase acceptance of the obtained triazole linkage in PCR.
中文翻译：
高效的DNA单击反应取代了酶促连接。
我们报告了基于铜催化的叠氮化物-炔烃环加成反应（CuAAC）的化学DNA-DNA连接方法。我们证明了离子的加入会极大地影响所谓的点击反应的效率。即使没有任何其他添加，例如通常用于预组织的夹板寡核苷酸，“点击连接”也可产生高达〜83％的产物，而没有任何副产物。另外，所需产物的纯化是直接的。与用于将衔接子引入例如mRNA文库制备中的酶促连接方法相比，这种靶向化学连接方法显示出几个优点：连接产物增加，并且没有衔接子或cDNA寡聚体副产物。
更新日期：2020-01-09
中文翻译：
高效的DNA单击反应取代了酶促连接。
我们报告了基于铜催化的叠氮化物-炔烃环加成反应（CuAAC）的化学DNA-DNA连接方法。我们证明了离子的加入会极大地影响所谓的点击反应的效率。即使没有任何其他添加，例如通常用于预组织的夹板寡核苷酸，“点击连接”也可产生高达〜83％的产物，而没有任何副产物。另外，所需产物的纯化是直接的。与用于将衔接子引入例如mRNA文库制备中的酶促连接方法相比，这种靶向化学连接方法显示出几个优点：连接产物增加，并且没有衔接子或cDNA寡聚体副产物。
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