DNA labeling for improved synthetic long reads

Project SummaryLong-read sequencing has the potential to greatly simplify sequencing, helping to accelerate scientists’ ability to perform de novo sequencing, haplotype phasing and transcriptomics. This project aims to develop a method to label DNA prior to next-generation sequencing, that maintains information about the proximity of fragments in the original strand, aiding in the downstream assembly of the sequencing data. To label the DNA, transposase will be loaded with specially-designed transposons containing barcode labels, and used to fragment the DNA, prior to next-generation sequencing. The first aim of this project is to construct the transposomes used to label the DNA. The second aim is to use these transposomes to tagment and sequence a model DNA system to demonstrate read lengths of ~50 kb using next-generation sequencing on less than a picogram of DNA. If successful, the sequencing approach developed in this grant will simplify synthetic long read sequencing, making high-accuracy and inexpensive long read sequencing more accessible to the genetics community.Project NarrativeBecause the genetic instructions contained in DNA play a crucial role in the healthy functioning of the body, it is often important for researchers and clinicians to read this information using DNA sequencing devices. To do this, the DNA is usually broken into small “words” that the sequencer can read, and then computer algorithms are used to guess the original order that those words appeared in the DNA, before it was broken apart. This project aims to develop a new way to molecularly label DNA fragments, to make it easier to determine the original DNA sequence.