A new enzymatic route for production of long 5'-phosphorylated oligonucleotides using suicide cassettes and rolling circle DNA synthesis |
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Authors: | Jakob S Lohmann Magnus Stougaard Jørn Koch |
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Institution: | (1) Institute of Pathology, Aarhus University, 8000 Aarhus C, Denmark |
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Abstract: | Background The quality of chemically synthesized oligonucleotides falls with the length of the oligonucleotide, not least due to depurinations
and premature termination during production. This limits the use of long oligonucleotides in assays where long high-quality
oligonucleotides are needed (e.g. padlock probes). Another problem with chemically synthesized oligonucleotides is that secondary
structures contained within an oligonucleotide reduce the efficiency of HPLC and/or PAGE purification. Additionally, ligation
of chemically synthesized oligonucleotides is less efficient than the ligation of enzymatically produced DNA molecules. |
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