HiFi sequencing of Hormonema macrosporum: Genome
A sequence of nucleotides incorporated by the DNA polymerase while reading a template, such as a circular SMRTbell (TM) template. Polymerase reads are most useful for quality control of the instrument run. Polymerase read metrics primarily reflect movie length and other run parameters rather than insert size distribution. Polymerase reads are trimmed to include only the high quality region; they include sequences from adapters; and can further include sequence from multiple passes around a circular template.Each polymerase read is partitioned to form one or more subreads, which contain sequence from a single pass of a polymerase on a single strand of an insert within a SMRTbell (TM) template and no adapter sequences. The subreads contain the full set of quality values and kinetic measurements. Subreads are useful for de novo assembly.The HiFi reads were generated using the circular consensus sequencing (CCS) application, tools included in SMRT Link. It has advantages to identify variants from single nucleotide to structural variant and to perform de novo assembly. In the case of Illumina data, the sequencing library is prepared by random fragmentation of the DNA or cDNA sample, followed by 5 and 3 adapter ligation. This library is loaded into a flow cell where fragments are captured on a lawn of surface-bound oligos complementary to the library adapters. As all 4 reversible, terminator-bound dNTPs are persent during each sequencing cycle, natural competition minimizes incorporation bias and greatly reduces raw error rates compared to other technologies. Then, sequencing data is converted into raw data for the analysis.