a new enzymatic route for production of long 5-phosphorylated oligonucleotides using suicide cassettes and rolling circle dna synthesis新的生产长5 磷酸化酶途径使用自杀式磁带和滚动循环dna合成寡核苷酸.pdfVIP

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a new enzymatic route for production of long 5-phosphorylated oligonucleotides using suicide cassettes and rolling circle dna synthesis新的生产长5 磷酸化酶途径使用自杀式磁带和滚动循环dna合成寡核苷酸.pdf

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a new enzymatic route for production of long 5-phosphorylated oligonucleotides using suicide cassettes and rolling circle dna synthesis新的生产长5 磷酸化酶途径使用自杀式磁带和滚动循环dna合成寡核苷酸

BMC Biotechnology BioMed Central Methodology article Open Access A new enzymatic route for production of long 5-phosphorylated oligonucleotides using suicide cassettes and rolling circle DNA synthesis Jakob S Lohmann, Magnus Stougaard and Jørn Koch* Address: Institute of Pathology, Aarhus University, 8000 Aarhus C, Denmark Email: Jakob S Lohmann - jlohm@as.aaa.dk; Magnus Stougaard - mstou@as.aaa.dk; Jørn Koch* - jokoc@as.aaa.dk * Corresponding author Published: 16 August 2007 Received: 28 March 2007 Accepted: 16 August 2007 BMC Biotechnology 2007, 7:49 doi:10.1186/1472-6750-7-49 This article is available from: /1472-6750/7/49 © 2007 Lohmann et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 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 synthesiz