Poster Presentation 40th Annual Lorne Genome Conference 2019

DNA Nanoballs and Single-Tube LFR: Affordable "Perfect" Genome Sequencing (#283)

Radoje Drmanac 1 , Xun Xu 2 , Brock A Peters 1
  1. Complete Genomics, MGI, San Jose, California, US
  2. BGI Research, BGI Shenzhen, Shenzhen, Guangdong, China

Precise personalized medicine depends on complete, accurate and phased WGS tests that are widely affordable. Our advanced massively parallel NGS platform based on the PCR-free error-free DNA nano-ball (DNB) nanoarrays (DNA spots <200nm), termed DNBseqTM, provides highly accurate pair-end 150-200 base WGS reads with extreme efficiency. DNB nanoarrays are prepared by rolling circle replication in solution generating ~300 copies of the original DNA template without making copy of a copy. DNB arrays are the most efficient patterned DNA arrays (<500nm pitch) enabling the lowest cost WGS.  In addition, DNB arrays provide several fold higher template density per spot resulting in higher quality sequencing data than PCR-based cluster arrays that also suffer from amplification errors. Recent improvements in making PCR-free WGS libraries and DNB arrays with ~500 base sequencing templates enable WGS with highest accuracy in detecting both SNPs and indels. These nanoarrays combined with novel fluidics and fast imaging cameras and scanning stages enable an order of magnitude more efficient generation of sequence data.  We expect to be the first to achieve $100 genome. Single-tube Long Fragment Read (stLFR) technology that generates uniquely co-barcoded reads for each 30kb-300kb long genomic DNA fragment at the cost of regular libraries is another revolutionary genomic technology we recently developed (Nature 2012, bioRxiv 2018). It allows separate (phased) assembly of parental chromosome sequences. Using up to 100 million barcodes from a pool of ~2 billion barcodes coupled to microbeads we uniquely barcode ~10 million long DNA fragments from 1ng of human DNA. Using co-barcoded reads, accurate WGS including detection and phasing of de novo mutations (~1 error per Gb, ~6 errors per genome) is achievable from ~10 cells (Genome Research 2015). In addition, stLFR’s uniquely co-barcoded reads obtained on DNBseqTM platform enable affordable “perfect” (de novo assembled accurate and phased) personal WGS.