A method for assembling synthetic long reads. James A. Stapleton Jeongwoon Kim John P. Hamilton Ming Wu Luiz C. Irber Rohan Maddamsetti Bryan Briney Linsey Newton Dennis R. Burton C. Titus Brown Christina Chan C. Robin Buell Timothy A. Whitehead 10.1371/journal.pone.0147229.g001 https://plos.figshare.com/articles/figure/_A_method_for_assembling_synthetic_long_reads_/1639540 <p>(a) Schematic of the approach. A supplemental barcode-pairing protocol (grey box) resolves the two distinct barcodes affixed to each original target molecule. (b) Reads associated with two distinct barcodes are shown aligned to the <i>E</i>. <i>coli</i> MG1655 reference genome. Barcode pairing merges the groups (bottom), increasing and evening the coverage and allowing assembly of the full 10-kb target sequence. (c) Length histogram of synthetic reads assembled from <i>E</i>. <i>coli</i> MG1655 genomic reads (minimum length 1 kb). The N50 length of the synthetic reads is 6.0 kb, and the longest synthetic read is 11.6 kb. (d) Mismatch rates of synthetic reads from the <i>E</i>. <i>coli</i> MG1655 dataset as a function of relative position along the synthetic read. (e) Length histogram of synthetic long reads assembled from <i>Gelsemium sempervirens</i> genomic reads (minimum length 1.5 kb). The N50 length of the synthetic reads is 4.3 kb. (f) An additional multiplexing index region (grey square) allows adapter-ligated samples to be mixed and processed in a single tube. Genomic DNA from twenty-four experimentally evolved strains of <i>E</i>. <i>coli</i> was separately ligated to adapters and amplified, then mixed into a single tube for the remaining steps of the protocol. <i>E</i>. <i>coli</i> genome coverage and N50 length are plotted for synthetic reads from each strain. Circle size indicates the number of short reads demultiplexed to a given strain.</p> 2016-01-28 12:38:22 cancer cell lines Long Reads acid molecules approach method mixture dna HIV env gene variants mRNA sequences animal genomic samples 11.6 kilobases length sequencing libraries custom equipment