Development & evaluation of Morphoseq synthetic long read sequencing technology

This project aims to evaluate a new method for generating synthetic long reads from short read (Illumina, BGI, etc) sequencing data. Long templates (8-10kbp) are randomly mutagenized using primer extension in the presence of the nucleotide analogue dPTP, which can base pair with A or G. The dPTP is then removed and a random set of mutations are fixed via a further primer extension in the presence of only natural nucleotides. Finally, a targeted number of the mutagenized templates are replicated with PCR and used to make a short read sequencing library. Short reads from the same template can be identified via the presence of shared mutations in overlapping regions.