Streamlined three-step workflow. Go from purified gDNA to sequencer-ready libraries in just 2.5 hours with the QIAseq FX DNA Library Kit.
The QIAseq FX DNA Library Kit generates customizable, reproducible DNA fragmentation. A Fragmentation of samples using the QIAseq FX DNA Library Kit is highly customizable. Three separate libraries were produced to demonstrate the wide range of fragment sizes possible using the QIAseq FX kit. Libraries shown include inserts approximately 200 (green), 500 (blue) or 1000 (red) bp long. Data were generated using an Agilent® Bioanalyzer.
DNA samples with broad GC contents were fragmented using the QIAseq FX DNA Library it with either a 5- or 10-minute run time. Fragment sizes were highly reproducible for all samples for each run time.
Due to its highly sequence-independent fragmentation, QIAseq FX exhibits minimal GC bias, which is comparable to mechanical shearing. 100 ng genomic DNA was used as the input in comparative library preparation and sequencing. The QIAseq FX DNA Library Kit and the leading mechanical shearing method (Supplier C) produced similar and consistent coverage across a wide range of GC contents. An all-enzymatic protocol from Supplier N yielded much higher bias toward high % GC. 1 ng genomic DNA was used as the input in a similar experiment. Again, QIAseq FX and mechanical shearing provided similar and consistent coverage, outperforming other methods.
Other supplier methods: Supplier C: mechanical shearing combined with a standard library prep; Supplier N: fragmentase; Supplier I: tagmentation.
For this comparative distribution experiment, the input was 100 ng samples of an equimolar mixture of genomic DNA from three bacterial species with vastly different GC contents: Fusobacterium nucleatum with 27% GC; Escherichia coli with 50% GC; and Bordetella pertussis with 67% GC (with the exception of the sample undergoing tagmentation, as only specific input amounts are accepted). The single, narrow peak for the library created with QIAseq FX DNA Library Kit shows that the majority of genomic targets have very similar total coverage depth. This is comparable to DNA fragmented by mechanical shearing.
Other supplier methods: Supplier C: mechanical shearing combined with standard library prep; Supplier N: fragmentase; Supplier I: tagmentation.
1 ng (all kits including QIAseq FX) or 100 ng (QIAseq FX only) genomic DNA was used as the input in a comparative duplication rate experiment. At 1 ng of input, the QIAseq FX DNA Library Kit performs comparably to mechanical shearing. At 100 ng input, QIAseq FX duplication rates are even lower, approaching the level typically seen from PCR-free workflows.
Other supplier methods: Supplier C: mechanical shearing combined with standard library prep; Supplier N: fragmentase; Supplier I: tagmentation.
With or without PCR, the QIAseq FX DNA Library Kit gives comparable genome coverage to mechanical shearing.
PCR-free library yields from the QIAseq FX DNA Library Kit are highly linear, with the amount of total library generated directly proportional to the amount of input DNA.
Other supplier methods: Supplier C: mechanical shearing combined with a standard library prep.
