Phage-free production of single-stranded DNA with user-defined sequence using Escherichia coli

Karl Behler, doctoral thesis Technical University of Munich, 2022

Appropriate distribution of the M13 phage genes between helper plasmid and phagemid enabled the scalable production of single-stranded DNA (ssDNA) with user-defined sequence in a fed-batch process with Escherichia coli in a stirred-tank bioreactor without making use of self-replicating phages. This process increased the maximum ssDNA concentration by a factor of 40 compared to literature data, which was also demonstrated on a m3 scale with an overall ssDNA isolation yield of 92%.


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