Single kb kb molecule sequencing to rapid and cost effectively assemble high quality arthropod genomes de novo

Nearly half the world s population is infected with at least one vector borne pathogen Many of these are carried and transmitted by arthropod vectors Despite huge economic and public health impacts of these arthropod vectors and efforts to sequence and assemble their genomes for the past decades the vector reference genomes are of low quality or non existent except the species with small genomes like mosquitoes High quality de novo genomic sequencing assembling and annotation of arthropod vectors are challenging due to size and complexity Novel high throughput sequencing technologies are urgently needed We propose to apply large DNA techniques to prepare high molecular weight HMW arthropod genomic DNA randomly shear the HMW DNA use superior enzyme reagents for the end repair A tailing nick gap fixing and adapter ligation of ultra length DNA construct unbiased MinION libraries of average kb and kb to prove the single molecule sequencing average kb and kb concept We ll demonstrate the feasibility of an advanced methodology pipeline for rapid high quality de novo genome assembly of three tick genomes e g Ixodes scapularis Dermacentor variabilis Amblyomma americanum with different genomic characteristics rt Gb genome sizes and diverse repetitive DNA to contiguity N rt Kb quality or completely finished genomes