Next Generation DNA Synthesis

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Ever large pieces of DNA such as genes and gene clusters are required for Synthetic Biology, and these are normally made by a combination of chemical and biochemical methods. The chemical methodology is required at the start of the process to generate very short pieces of DNA (oligonucleotides) by automated solid-phase methods. These are then used to build bigger pieces of DNA by biochemical methods that are based on the polymerase chain reaction (PCR amplification). The chemical synthesis of DNA can lead to damage which results in mistakes (mutations) in the final DNA product, and to avoid this the DNA has to be repaired by various enzymes. This is tedious and slows down the overall process, increasing costs and limiting the size of DNA that can be made. In this project we will analyse DNA made by modern ultra high throughput chemical methods and optimise the process to minimise mutations. We will also explore a different way to make large pieces of DNA; enzymatic ligation. In this process DNA constructs with modified bases can be made, which are very useful in gene expression and biomedical studies. These cannot be made by PCR amplification which erazes the modifications. Such modified DNA can only be properly made from highly pure oligonucleotides in very large numbers, placing stringent requiremenst on high-throughput oligonucleotide synthesis. Overall this project will greatly increase the capacity, quality and efficiency of DNA synthesis and is highly relevant to Synthetic Biology Centres in the UK and beyond.