ABSTRACT
Next generation sequencing (NGS) produces large quantities of data – exponentially more than traditional methods for STR typing and single nucleotide polymorphism (SNP) analysis. Software is used to interpret the fluorescence emission or voltage changes and generate a raw Deoxyribonucleic acid (DNA) sequence for each cluster. The number of sequences of each type is counted, and tables are generated showing the number of “reads” or counts of each sequence recorded. Phasing is detected upon comparing the subsequent base sequence addition to other clusters. The Illumina MiSeq and the Verogen MiSeq FGx record a photographic image of all of the fluorescing clusters on the flow cell after the addition of the base in each cycle. The MiSeq FGx outputs metrics for cluster density, percent of clusters passing the filter, and phasing and prephasing that indicate the quality of the run and issue warning flags when the values are outside the set range.
